Tricyclic compound, pharmaceutical composition and application

Abstract

The invention discloses a tricyclic compound, a pharmaceutical composition and application, the structure of the compound is shown as a formula I or a formula II, and the tricyclic compound further comprises an isomer, pharmaceutically acceptable salt or a mixture of the isomer and the pharmaceutically acceptable salt. The compound and the pharmaceutical composition thereof can effectively inhibit the respiratory syncytial virus, can be prepared into a drug for treating respiratory syncytial virus infection, can exert a drug effect at a molecular level, has a more excellent treatment effect, and can optimally reach a nanomolar concentration level. In addition, the compound preparation method is simple and easy to operate. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a tricyclic compound, a pharmaceutical composition and an application. Background Art

[0002] Respiratory syncytial virus (RSV) was first discovered and isolated from chimpanzees with respiratory diseases in 1955. In 1957, RSV was isolated from infants with severe lower respiratory tract diseases. Because the virus can cause fusion lesions in cells, it was named human respiratory syncytial virus (hRSV). RSV infection is widely prevalent around the world. The RSV epidemic season has obvious climatic distribution characteristics. In temperate regions, the RSV epidemic period generally lasts for 2 to 5 months, and the peak usually occurs in winter. In tropical regions, syncytial virus lasts longer than in temperate regions, and the peak usually coincides with the rainy season. The clinical mild symptoms of RSV infection are respiratory tract infections, which can be manifested as cough, rhinitis, occasional fever, or severe lower respiratory tract infections leading to bronchiolitis, pneumonia or acute respiratory failure. RSV infection seriously endangers human life and health.

[0003] Currently, only ribavirin and palivizumab are approved for the treatment of RSV infection. Ribavirin has poor safety, while palivizumab is expensive, which limits its clinical application. Therefore, the development of economical, safe and effective RSV small molecule drugs has broad market prospects and social benefits. Summary of the invention

[0004] Object of the Invention: In response to problems existing in the prior art, the present invention provides a tricyclic compound, which or a pharmaceutically acceptable salt thereof has high RSV inhibitory activity.

[0005] The present invention also provides pharmaceutical compositions of the tricyclic compounds and their applications.

[0006] Technical Solution: To achieve the above-mentioned object, a tricyclic compound or a pharmaceutically acceptable salt thereof according to the invention has the structure of formula I or formula II:

[0007]

[0008] Wherein, the three-ring segment ring I, ring II or ring III are each independently selected from C5-C 20 Aromatic ring, C5-C 20 Aromatic heterocycle, C5-C 20 Substituted aromatic ring or C5-C 20Substituted aromatic heterocycle, the C5-C 20 The substituted aromatic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, respectively, and the C5-C 20 Substituted aromatic heterocycles are C5-C 20 The aromatic heterocycles are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, respectively, and the C5-C 20 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0009] As preferred, the tricyclic segment ring I, ring II or ring III are each independently selected from C5-C 15 Aromatic ring, C5-C 15 Aromatic heterocycle, C5-C 15 Substituted aromatic ring or C5-C 15 Substituted aromatic heterocycle, the C5-C 15 The substituted aromatic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, respectively, and the C5-C 15 Substituted aromatic heterocycles are C5-C 15 The aromatic heterocycles are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, respectively, and the C5-C 15 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0010] As more preferred, the tricyclic segment ring I, ring II or ring III are each independently selected from the group consisting of C5-C 10 Aromatic ring, C5-C 10 Aromatic heterocycle, C5-C 10 Substituted aromatic ring or C5-C 10 Substituted aromatic heterocycle, the C5-C 10 The substituted aromatic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, respectively, and the C5-C 10 Substituted aromatic heterocycles are C5-C 15 The aromatic heterocycles are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, respectively, and the C5-C 10 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0011] Ring A is C5-C 15 Aliphatic ring, C5-C 15 Spirocyclic, C5-C 15 Heterospirocyclic, C5-C 15 Bridge ring, C5-C 15 Heterobridged ring, C5-C 15 Aliphatic heterocyclic, C5-C 15 Aromatic ring, substituted C5-C15 Aliphatic ring or substituted C5-C 15 Aromatic ring, the C5-C 15 Heterospirocyclic, C5-C 15 Heterobridged ring or C5-C 15 Fat heterocyclic ring contains 1 to 2 independent N atoms, the C5-C 15 The aromatic heterocycle contains 1 to 3 independently selected from N or O atoms, the substituted C5-C 15 Aliphatic ring and C5-C 15 The aromatic rings are optionally substituted with 1 to 2 individually selected from -CF3 or halogen substituents;

[0012] Preferably, ring A is C5-C 12 Aliphatic ring, C5-C 12 Spirocyclic, C5-C 12 Heterospirocyclic, C5-C 12 Bridge ring, C5-C 12 Heterobridged ring, C5-C 12 Aliphatic heterocyclic, C5-C 12 Aromatic ring, substituted C5-C 12 Aliphatic ring or substituted C5-C 12 Aromatic ring, the C5-C 12 Heterospirocyclic, C5-C 12 Heterobridged ring or C5-C 12 Fat heterocyclic ring contains 1 to 2 independent N atoms, the C5-C 12 The aromatic heterocycle contains 1 to 3 independently selected from N or O atoms, the substituted C5-C 12 Aliphatic ring and C5-C 12 The aromatic rings are optionally substituted with 1 to 2 individually selected from -CF3 or halogen substituents;

[0013] More preferably, ring A is C5-C 10 Aliphatic ring, C5-C 10 Aliphatic heterocyclic, substituted C5-C 10 Aliphatic ring or substituted C5-C 10 Aliphatic heterocyclic ring, the C5-C 10 Aliphatic heterocyclic or substituted C5-C 10 Fat heterocyclic ring contains 1 to 2 independent N atoms, the substituted C5-C 10 Aliphatic ring and C5-C 10 The fat heterocycles are optionally substituted with 1 to 2 individually selected from -CF3 or halogen substituents;

[0014] Ring B is C3-C 12 Aliphatic ring, C3-C 12 Spirocyclic, C3-C 12 Bridge ring, C3-C 12Aliphatic heterocyclic, C5-C 12 Aromatic ring, C3-C 12 Aromatic heterocycle, substituted C3-C 12 Aliphatic ring, substituted C5-C 12 Aromatic ring or substituted C3-C 12 Aromatic heterocyclic ring, the C3-C 12 Aliphatic heterocyclic ring or C3-C 12 The aromatic heterocycle contains 1 to 3 independently selected from N or O atoms, the substituted C3-C 12 Aliphatic ring or substituted C5-C 12 Aromatic ring or C3-C 12 The aromatic heterocycles are optionally substituted by 0 to 2 substituents independently selected from -CH3, -OCH3, -F, or -CF3, respectively, and the C3-C 12 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0015] Preferably, ring B is C4-C 11 Aliphatic ring, C4-C 11 Spirocyclic, C4-C 11 Bridge ring, C4-C 11 Aliphatic heterocyclic, C4-C 11 Aromatic ring, C4-C 11 Aromatic heterocycle, substituted C4-C 11 Aliphatic ring, substituted C4-C 11 Aromatic ring or substituted C4-C 11 Aromatic heterocyclic ring, the C4-C 11 Aliphatic heterocyclic or C4-C 11 The aromatic heterocycle contains 1 to 3 independently selected from N or O atoms, the substituted C4-C 11 Aliphatic ring or substituted C4-C 11 Aromatic ring or C4-C 11 The aromatic heterocycles are optionally substituted by 0 to 2 substituents independently selected from -CH3, -OCH3, -F, or -CF3, respectively, and the C4-C 11 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0016] More preferably, ring B is a C4-C6 fatty heterocycle and C6-C 10 The spiral ring, C7-C8 bridge ring, C4-C8 aromatic ring, replaces the C4-C6 fatty ring or replaces the C4-C8 aromatic ring, the C4-C6 fatty heterocycle contains 1 to 2 independently selected from N atoms, and the substituted C4-C 11 The aromatic rings are optionally substituted with 0 to 2 substituents independently selected from -CH3, -OCH3, -F, or -CF3, respectively;

[0017] R 1 ,R2 ,R 3 ,R 4 ,R 5 Each is independently selected from -H, -D, halogen, -CH3, -C2H5, -CF3, -SF5, -CD3, C3-C4 cycloalkyl, -OCH3, -OCF3, -NH2, -NO2 or -CN;

[0018] X is -C-, -N or -NO-;

[0019] Q is -C- or -N-, where when Q is -C-, Q is the chiral center, preferably in the R configuration;

[0020] Y, Z are each independently selected from -CH2-, -NH-, -N(CH3)-, -O-, -S-, -SO- or -SO2-;

[0021] The chemical bond between Y and Z is a single bond or a double bond, preferably a single bond;

[0022] The chemical bond between Q and A ring is a single bond or a double bond, preferably a single bond;

[0023] T is any integer in -O-, -NH-, -CO-, -CHF-, -CF2- or -(CH2)n-, n=0-4;

[0024] E is selected from -O-, -NH-, -CO-, -CHF-, -CF2-, -C(CH3)2-, or

[0025] -(CH2)n-, n=any integer from 0 to 4;

[0026] L is selected from -O-, -NH-, -CO-, -SO-, -SO2-, or -(CH2) n -, n = any integer from 0 to 4;

[0027] Preferably, L is selected from -O-, -NH-, or -(CH2) n -, n = any integer from 0 to 4;

[0028] More preferably, L is selected from -NH-, -CO-, -SO2- or -(CH2) n -, n = any integer from 0 to 4;

[0029] M is selected from -H, -D, -CH3, -CD3, -CF3, -OH, -NHR 6 ,-CONHR 6 、-SO2CH3、-CH2=NR 6、-(CH2) n -, any integer in n=0-4, C3-C6 fatty ring, C3-C6 fatty heterocycle, C5-C 12 Aromatic ring or C5-C 12 Aromatic heterocycle, the C3-C6 aliphatic heterocycle or C5-C 12 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0030] Preferably, M is selected from -H, -D, -CH3, -CD3, -CF3, -OH, -NHR 6 ,-CONHR 6 、-SO2CH3、-CH2=NR 6 、-(CH2) n -, any integer in n=0-4, C3-C6 fatty ring, C3-C6 fatty heterocycle, C5-C 10 Aromatic ring or C5-C 10 Aromatic heterocycle, the C3-C6 aliphatic heterocycle or C5-C 10 The aromatic heterocycle contains 1 to 3 heteroatoms independently selected from N, O or S;

[0031] More preferably, M is selected from -CH3, -CD3, -OH, -NHR 6 ,-CONHR 6 、-SO2CH3、-CH2=NR 6 、-(CH2) n -, any integer in n=0-4, C3-C6 fatty ring or C5-C6 aromatic ring, the C5-C6 aromatic heterocycle contains 1 to 2 independent N atoms;

[0032] R 6 Selected from -H, -D, -OH or -OCH3.

[0033] Preferably, the tricyclic fragment ring I, ring II or ring III are each independently selected from any one of the following groups:

[0034]

[0035] Preferably, the ring A is selected from any of the following groups:

[0036]

[0037] Preferably, the ring B is selected from any of the following groups:

[0038]

[0039] Preferably, the R 1 ,R2 ,R 3 Each independently selected from -H, -D, -F, -Cl or -Br; R 4 ,R 5 Each is independently selected from -H, -D, -F, -CH3, -CF3, -OCH3 or -OCF3.

[0040] Preferably, the structure of the -LM is such as Formula III or Formula IV:

[0041]

[0042] Where V is selected from -O-, -NH-, -CH2-, -CO-, -SO- or -SO2-, n=0-4;

[0043] U is selected from -CH-, -CH2-, or n = any integer between 0 and 4;

[0044] R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3, -CF3-, -OCH3, -OCF3, -COOH, -CONH2, -SO2NH2, -SO2CH3, -isopropyl, -cyclopropyl, -cyclohexyl or The R 9 Selected from -H, -OH, -NH2, -OCH3, -CH3, -CD3, -CF3-, -OCF3 or halogen;

[0045] R 8 Selected from -H, -CH3 or -CD3.

[0046] As more preferred, V is selected from any integer in -CH2-, -CO-, -SO- or -SO2-, n=0-3;

[0047] U is selected from -CH-, -CH2-, n = any integer between 0 and 3;

[0048] R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3, -CONH2, -isopropyl, -cyclopropyl, -cyclohexyl or The R 9 Selected from -H, -OH, -NH2, -OCH3, -CH3, -CD3, -CF3-, -OCF3 or halogen;

[0049] R 8 Selected from -H or -CH3.

[0050] As more preferred, V is selected from any integer in -CH2-, -CO-, n=1-2;

[0051] U is selected from -CH-, -CH2-, n = any integer between 0 and 2;

[0052] R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3, -CONH2, -isopropyl, -cyclopropyl or -cyclohexyl;

[0053] R 8 Selected from -H or -CH3.

[0054] As a more preferred embodiment, wherein V is selected from -CO-, n=1;

[0055] U is selected from any integer in -CH- or -CH2-, n=0-2;

[0056] R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3 or -CONH2;

[0057] R 8 Selected from -H or -CH3.

[0058] As the preferred tricyclic compound or a pharmaceutically acceptable salt thereof, it is selected from any of the following compounds:

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065] A pharmaceutical composition according to the invention comprises any of the compounds or pharmaceutically acceptable salts, stereoisomers, solvates, hydrates and pharmaceutically acceptable carriers thereof.

[0066] Wherein, the pharmaceutically acceptable salt is a salt formed by the compound and the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, carbonic acid, nitric acid, hydrobromic acid, hydroiodic acid, maleic acid, fumaric acid, tartaric acid, citric acid, malic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, succinic acid, acetic acid, mandelic acid, isobutyric acid or malonic acid.

[0067] Furthermore, the compound and a pharmaceutically acceptable carrier form a pharmaceutical composition, which is made into a common pharmaceutical preparation, such as tablets, capsules, syrups, suspensions or injections, and the preparation can be added with common pharmaceutical excipients such as flavorings, sweeteners, liquid / solid fillers, diluents, etc. The use of the compound or pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present invention in the preparation of a drug for preventing or treating viral infection.

[0068] Among them, the virus infection is respiratory syncytial virus infection.

[0069] The above-mentioned compounds or pharmaceutical compositions thereof of the present invention can be prepared as RSV inhibitor drugs, specifically for the treatment of respiratory syncytial virus infection.

[0070] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0071] (1) Such compounds and their pharmaceutical compositions can effectively inhibit RSV proliferation and inhibit EC at the cellular level. 50 The values ​​were mostly less than 200 nM, and the optimal value was less than 5 nM;

[0072] (2) Such compounds and their pharmaceutical compositions are widely used and can be prepared as drugs for treating RSV infection. They can exert their efficacy at the cellular and animal levels, and the therapeutic effects are even better, reaching nanomolar concentration levels;

[0073] (3) The compounds of the present invention are easy to prepare and are cheap, have simple structure, ingenious design, cheap and easy to obtain, safe synthesis process, environmentally friendly, and easy to produce on a large scale. . BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1Figure 4 is the in vivo efficacy of RSV infection in BALB / c mice. a. Schematic diagram of infection and treatment of BALB / c mice. b. Mice infected with RSVA2 were killed on day 4 after infection to detect infectious viral RNA in lung tissue using FFA (n=6). c. Mice infected with RSVA2 were killed on day 4 after infection to detect infectious viral RNA in lung tissue using FFA (n=6). d. Histological analysis, virus-induced pathology in the lungs of mice infected with RSVA2 was observed on day 4 after infection. e. Quantitative scoring of lung pathology in mice on day 4 after infection The histopathological score of each lung inflammation was 0-4, where 0 is a normal healthy lung and 4 is a severely pathological confluent area. These data are representative of at least two experiments. Error bars are SEM±mean. Statistical differences were determined by two-way ANOVA in b, c, and e. *P<0.05, **P<0.01, ***P<0.001; NS, not significant. DETAILED DESCRIPTION

[0075] The technical scheme of the present invention is further described below with reference to the examples. The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from companies such as Leyan, Bid Pharmaceuticals, Aladdin and Anaiji.

[0076] Example 1: Synthesis of 4-(2-(4-(dibenzo[b,e]oxazo-11(6H)-subunit)piperidin-1-yl)-2-oxoethyl)piperidin-1-formamide (1)

[0077]

[0078] Synthesis of intermediate 1-2:

[0079] Zn powder (10.00 g, 154 mmol) was dispersed in anhydrous THF (100 mL), added to a three-necked bottle. After N2 was replaced, the ice-salt bath was cooled to 10°C, and TiCl4 (5.75 mL, 52.3 mmol) was added dropwise to keep the temperature not exceeding 10°C. After the addition was completed, the temperature was raised to 80°C and reacted for 2h, followed by the addition of a mixed solution of dibenzo[b,e]oxazepine-11(6H)-one (5.00g, 23.8mmol, cas: 4504-87-4) and tert-butyl 4-oxopiperidine-1-carboxylate (5.70g, 28.5mmol, cas: 79099-07-3) in anhydrous THF (100mL), and the reaction was continued at 80°C for 2h. After the reaction was completed as monitored by TLC, the temperature was lowered, and then a saturated aqueous solution of K2CO3 was added to adjust the pH to 8. 50mL of ethyl acetate EA was added, followed by suction filtration, separation, and extraction of the filtrate with EA (50mL x 2). The organic layers were combined, washed once with water and once with saturated brine, concentrated in vacuo, and purified by column chromatography (DCM:MeOH=10:1) to obtain intermediate 1-2. 1 HNMR(300MHz, CDCl3)δ=7.49(s,1H),7.33–7.24(m,4H),7.20(s,1H),7.20–7.13(m,1H),6.99(s,1H),5.17 (d,J=1.0Hz,2H),2.85–2.77(m,4H),2.55(t,J=3.9Hz,4H),1.85–1.79(m,1H)ppm.HR-MS(ESI):Calculated for C 19 H 19 NO[M+H] + :278.1647, found 278.1632. Yield 62%.

[0080] Synthesis of intermediate 1-3:

[0081] 2-(1-(tert-Butyloxycarbonyl)piperidin-4-yl)acetic acid (0.53 g, 2.2 mmol) was added to a three-necked flask, and N,N-dimethylformamide DMF (80 mL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride EDCI (0.69 g, 3.6 mmol), 1-hydroxybenzotriazole HOBT (0.49 g, 3.6 mmol), triethylamine TEA (0.55 g, 5.4 mmol) were added, and stirred at room temperature for 30 min, and then intermediate 1-2 (0.5 g, 1.8 mmol) was added, and the reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water, and a solid precipitated, which was then filtered off with suction. After the filter cake was dried, it was purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain 0.65 g of intermediate 1-3.

[0082] 1 H NMR (300MHz, CDCl3) δ=7.49(s,1H),7.33–7.24(m,4H),7.20(s,1H),7.20–7.13(m,1H),6.99(s,1H),5.17(d,J=0.9H z,2H),3.66–3.60(m,1H),3.63–3.57(m,3H),3.43(s,2H),3.33–3.29(m,2H),2.63–2.52(m,4H),2.32(s,1H),2.24(s 1H),2.06–1.97(m,1H),1.9–1.83(m,4H)ppm.HR-MS(ESI):Calculated for C 31 H 38 N2O4[M+H] + :503.2832, found 503.2822. Yield 72%.

[0083] Synthesis of intermediate 1-4:

[0084] Intermediate 1-3 (0.2g) was added to a single-mouth bottle, EA (5mL) was added, 4NHCl-EA solution (2mL) was added under an ice bath, and then the reaction was carried out at room temperature for 2 hours. After TLC monitoring reaction was complete, intermediate 1-4 and 0.14g were obtained.

[0085] 1 H NMR(300MHz, CDCl3)δ=7.49(s,1H),7.34–7.24(m,4H),7.22–7.18(m,1H),7.19–7.1 7(m,1H),6.99(s,1H),5.17(d,J=0.9Hz,2H),3.74–3.67(m,2H),3.43(s,2H),2.93–2 .88(m,2H),2.78–2.74(m,2H),2.63–2.52(m,4H),2.37–2.28(m,2H),2.23(s,1H),1 .94–1.90(m,1H),1.59–1.53(m,2H),1.40–1.35(m,2H)ppm.HR-MS(ESI):Calculated forC 19 H 26 BN2O4[M+H] + :403.2307, found403.2311. Yield 88%.

[0086] Synthesis of 4-(2-(4-(dibenzo[b,e]oxazo-11(6H)-subunit)piperidin-1-yl)-2-oxoethyl)piperidin-1-formamide (1):

[0087] The intermediate 1-4 (0.1 mmol) was added to a single-mouth bottle, and a mixed solvent of N-methyl-2-pyrrolidone NMP:H2O (4:1, 15 mL) was added, followed by urea (0.1 mmol), and the mixture was refluxed at 130°C for 9 h. After the TLC reaction was complete, the temperature was lowered to 25°C, and the reaction solution was poured into water. Solids precipitated, which were then filtered and purified by column chromatography (DCM:MeOH=40:1) to obtain the final product 1.

[0088] 1 H NMR(300MHz, CDCl3)δ=7.49(s,1H),7.34–7.24(m,4H),7.24–7.17(m,1H),7.19–7 .17(m,1H),6.99(s,1H),5.29(s,2H),5.17(d,J=0.9Hz,2H),3.74–3.67(m,2H),3 .65–3.61(m,1H),3.61–3.57(m,2H),3.43(s,2H),2.63–2.52(m,4H),2.33(s,1H) ,2.25(s,1H),2.07–1.97(m,1H),1.88–1.74(m,4H).ppm.HR-MS(ESI):Calculated for C 26 H 30 BN4O4[M+H] + :473.2360, found 473.2357. Yield 68%.

[0089] Using the method in Example 1, dibenzo[b,e]oxazepine-11(6H)-one was replaced with 8-chloro-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridine-11-one (cas: 31251-41-9), and tert-butyl 4-oxopiperidine-1-carboxylate was replaced with N-Boc-nortropine (cas: 185099-67-6) to obtain compound 3:

[0090]

[0091] 1H NMR (300MHz, CDCl3) δ = 8.42 (s, 1H), 7.68 (s, 1H), 7.35 (s, 1H), 7.28 (s, 1H) ,7.20–7.12(m,2H),5.29(s,2H),4.14(s,2H),3.71–3.63(m,4H),3.29–3.1 6(m,2H),2.88(s,1H),2.87–2.81(m,1H),2.79–2.69(m,4H),2.28(s,1H),2 .21(s,1H),2.05–1.92(m,3H),1.88–1.67(m,6H).HR-MS(ESI):Calculated for C 29 H 33 ClN4O2[M+H] + :505.2395,found505.2393.

[0092] Using the method in Example 1, dibenzo[b,e]oxazepine-11(6H)-one was replaced with 8-chloro-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridine-11-one (cas: 31251-41-9), and 4-oxopiperidine-1-carboxylic acid tert-butyl ester was replaced with 6-oxo-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (cas: 1181816-12-5) to obtain compound 5:

[0093]

[0094] 1 H NMR (300MHz, CDCl3) δ = 8.42 (s, 1H), 7.68 (s, 1H), 7.35 (S, 1H), 7.28 (s, 1H), 7.20–7.12 (m, 2H),5.29(s,2H),3.76(s,2H),3.74–3.67(m,1H),3.70–3.66(m,1H),3.65(s,2H),3.66–3. 61(m,1H),3.61(s,1H),3.29–3.16(m,2H),2.93–2.85(m,1H),2.87–2.81(m,1H),2.80(s, 3H),2.28(d,J=7.0Hz,2H),2.06–1.97(m,1H),1.88–1.74(m,4H).HR-MS(ESI):Calculated for C 28 H 31 ClN4O2[M+H] + :491.2208,found 491.2205.

[0095] Using the synthesis method of compound 1 in Example 1, dibenzo[b,e]oxazo-11(6H)-one was replaced with 6,11-dihydrodobenzo[b,e]thiazo-11-one (cas: 1531-77-7) to obtain compound 8:

[0096]

[0097] 1 H NMR(300MHz, CDCl3)δ=7.49(s,1H),7.34–7.24(m,4H),7.24–7.17(m,1H),7.19– 7.17(m,1H),6.99(s,1H),5.29(s,2H),5.17(d,J=0.9Hz,2H),3.74–3.67(m,2H) ,3.65–3.61(m,1H),3.61–3.57(m,2H),3.35(s,2H),2.63–2.52(m,4H),2.33(s, 1H),2.25(s,1H),2.07–1.97(m,1H),1.88–1.74(m,4H).HR-MS(ESI):Calculated for C 27 H 31 N3O2S[M+H] + :462.2210, found 462.2200.

[0098] Using the synthesis method of compound 1 in Example 1, dibenzo[b,e]oxazo-11(6H)-one was replaced with thiaxon-9-one (cas: 492-22-8) to obtain compound 9:

[0099]

[0100] 1 H NMR(300MHz, CDCl3)δ=7.67–7.60(m,1H),7.54–7.43(m,2H),7.41–7.35(m,1H),5.29(s,1H),3.69–3.57(m,1H),3.66–3.57 (m,2H),3.42(s,1H),2.63–2.52(m,2H),2.37–2.21(m,1H),2.06–1.97(m,1H),1.88–1.74(m,2H).HR-MS(ESI):Calculated forC 26 H 29 N3O2S[M+H] + :448.2053, found 448.2050.

[0101] The synthesis method of compound 1 in Example 1 was used to replace dibenzo[b,e]oxazepine-11(6H)-one with 3-chloro-6-methyldibenzo[c,f][1,2]thiazepine-11(6H)-one 5,5-dioxide (cas: 26638-53-9) to obtain compound 10:

[0102]

[0103] 1 H NMR (300MHz, CDCl3) δ = 7.75 (d, J = 2.2Hz, 1H), 7.53 (d, J = 8.6Hz, 1H), 7.49 (s, 1H), 7.40 (s ,1H),7.35–7.25(m,2H),7.25–7.18(m,1H),5.29(s,2H),3.74–3.67(m,1H),3.70–3.66(m ,1H),3.66–3.61(m,1H),3.64–3.57(m,3H),3.43(s,2H),3.41(s,3H),2.63–2.52(m,4H), 2.33(s,1H),2.25(s,1H),2.06–1.96(m,1H),1.88–1.74(m,4H).HR-MS(ESI):Calculated for C 27 H 31 ClN4O4S[M+H] + :543.1827, found 543.1825.

[0104] Example 2: Synthesis of 2-(3-aminopyrrolidin-1-yl)-1-(4-(dibenzo[b,e]oxazo-11(6H)-subunit)piperidin-1-yl)e-1-one (2)

[0105]

[0106] Synthesis of intermediate 2-1:

[0107] The intermediate 1-2 (0.5 g, 1.80 mmol) was added to a single-mouth bottle and dissolved with 5 mL of dichloromethane (DCM). Chloroacetyl chloride (0.24 g, 2.16 mmol) and TEA (0.37 g, 3.61 mmol) were added under ice bath, and then reacted at room temperature for 2 h. After the reaction was complete by TLC, saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted with DCM three times. The organic phases were combined, dried, and concentrated to obtain the intermediate 2-1, which was directly used for the next step without purification.

[0108] Synthesis of intermediate 2-2:

[0109] The intermediate 2-1 (0.30 g, 0.84 mmol) was added to a single-necked bottle, dissolved with 5 mL of CH3CN, and tert-butyl pyrrolidine-3-carbamate (0.19 g, 1.02 mmol) and K2CO3 (0.24 g, 1.7 mmol) were added, followed by reaction at 75°C for 2 h. After the TLC reaction was complete, it was diluted with water, extracted with EA three times, and the organic phases were combined, dried, and concentrated to obtain the intermediate 2-2, which was directly used for the next step without purification.

[0110] Synthesis of 2-(3-aminopyrrolidin-1-yl)-1-(4-(dibenzo[b,e]oxazo-11(6H)-subunit)piperidin-1-yl)e-1-one (2):

[0111] The intermediate 2-2 (0.43 g, 0.84 mmol) was added to a single-necked bottle, dissolved with 10 mL of EA, and 3 mL of 4N HCl-EA solution was added in an ice bath, followed by reaction at room temperature for 1 h. After the reaction was complete by TLC, the mixture was filtered, slurried with EA, and dried to obtain 20.35 g of the compound. 1 H NMR(300MHz, CDCl3)δ=7.49(s,1H),7.34–7.24(m,4H),7.25–7.18(m,1H),7.17–2.1 2(m,1H),6.99s,1H),5.17(d,J=0.9Hz,2H),3.60(s,2H),3.49(s,2H),3.27–3.20(m, 1H),3.13(d,J=4.9Hz,2H),2.89–2.83(m,1H),2.81–2.72(m,2H),2.62(s,1H),2.62 –2.52(m,4H),2.22(s,1H),2.09(s,1H),1.73–1.57(m,2H).HR-MS(ESI):Calculated forC 25 H 29 N3O2[M+H] + :404.2333 found 404.2330. Yield 85%.

[0112] The synthesis method of compound 1 in Example 1 and the synthesis method of compound 2 in Example 2 were used to replace dibenzo[b,e]oxazepine-11(6H)-one with 8-chloro-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridine-11-one (cas: 31251-41-9) to obtain compound 4:

[0113]

[0114] 1H NMR (300MHz, CDCl3) δ = 8.42 (s, 1H), 7.68 (s, 1H), 7.35 (dd, J = 7.9, 4.4Hz, 1H), 7.28 (s, 1H), 7.20–7 .12(m,2H),4.19–4.12(m,2H),3.29–3.17(m,3H),3.16(d,J=3.7Hz,2H),2.94–2.81(m,3H),2.81–2 .74(m,1H),2.78–2.69(m,5H),2.64–2.58(m,1H),2.22(s,1H),2.09(s,1H),2.06–1.93(m,2H),1. 83–1.76(m,1H),1.79–1.72(m,1H),1.76–1.69(m,1H),1.66–1.57(m,1H).HR-MS(ESI):Calculated for C 27 H 31 ClN4O[M+H] + :463.2259,found463.2255.

[0115] The synthesis method of compound 1 in Example 1 and the synthesis method of compound 2 in Example 2 were used to replace dibenzo[b,e]oxazepine-11(6H)-one with 8-chloro-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridine-11-one (cas: 31251-41-9), and tert-butyl 4-oxopiperidine-1-carboxylate with tert-butyl 6-oxopiperidine-2-carboxylate (cas: 1181816-12-5) to obtain compound 6:

[0116]

[0117] 1 H NMR (300MHz, CDCl3) δ = 8.42 (s, 1H), 7.68 (s, 1H), 7.35 (s, 1H), 7.28 (s, 1H), 7.17 (d,J=8.9Hz,1H),7.14(s,1H),3.75(s,2H),3.65(s,2H),3.29–3.16(m,4H),3.1 4(d,J=13.0Hz,1H),2.94–2.71(m,6H),2.80(s,4H),2.64–2.58(m,1H),2.22(s, 1H),2.09(s,1H),1.83–1.74(m,1H),1.65–1.60(m,1H).HR-MS(ESI):Calculated for C 26 H 29ClN4O[M+H] + :449.2103, found 449.2105.

[0118] Example 3: Synthesis of 1-(2-(4-(3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptane[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidin-4-carboxamide (14)

[0119]

[0120] Synthesis of Intermediate II:

[0121] I (5-bromo-2-cyanopyridine) (25 g, 126.9 mmol, cas: 97483-77-7) was added to 80 mL of tert-butyl alcohol, and the temperature was raised to 70 ° C until the raw materials were completely dissolved. 20 mL of 98% concentrated sulfuric acid was slowly added to the reaction solution. After 10 min, TLC monitored the reaction to be complete. The reaction solution was cooled to room temperature and slowly poured into ice water. Ammonia water was used to adjust the pH to 7, and ethyl acetate was extracted three times. The organic layers were combined and washed with water twice, washed with saturated brine once, and dried over anhydrous sodium sulfate and evaporated under reduced pressure to remove the solvent to obtain 37 g of intermediate II as a yellow oil, which was directly used in the next step without purification.

[0122] Synthesis of intermediate III:

[0123] The intermediate II (37g, 136.45mmol) was dissolved in dry THF (400mL), and the temperature was lowered to -60°C after nitrogen replacement 3 times. Diisopropylamine lithium (140mL, 280mmol) was slowly added to the reaction solution, and m-chlorobenzyl chloride (27g, 163.74mmol) was added to the reaction solution after stirring at -60°C for 40min. After the addition was completed, the reaction was reacted at -60°C for 1h and then moved to room temperature to continue the reaction for 4h. The reaction was monitored by TLC. The reaction was quenched with saturated ammonium chloride aqueous solution, the organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate. The organic layers were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain 70g of intermediate III brown oil. The pure product was 31.4g of yellow oil by column chromatography (PE:EA=60:1), with a yield of 58%.

[0124] Synthesis of Intermediate IV:

[0125] Add 31.4 g of intermediate III to 150 mL of phosphorus oxychloride, heat to 110°C and reflux for 4 h. Monitor the reaction by TLC until complete. Cool the reaction solution to room temperature and slowly pour it into ice water. A grayish white solid precipitates and filter to obtain 20 g of intermediate IV. Use it directly in the next step without purification.

[0126] Synthesis of intermediate V:

[0127] Dissolve 20g of intermediate IV in 120mL of trifluoromethanesulfonic acid, heat to 60℃ for 2h, then cool to room temperature for 4h, add 6mol / L HCl solution (60mL) and heat to 110℃ for 2h. After TLC monitoring, cool to room temperature, pour the reaction solution into ice water, adjust to neutral with ammonia water, and filter to obtain a crude product. Column chromatography (PE:EA=60:1) gives white solid intermediate V13g.

[0128] Synthesis of intermediate V-1:

[0129] Intermediate V (13.01 g, 40.33 mmol) was dissolved in 65 mL of 98% concentrated sulfuric acid, cooled to 0°C, and concentrated nitric acid (5.85 mL, 2.3 eq) was slowly added. After 1.5 h, the reaction was complete as monitored by TLC. The reaction solution was poured into ice water, and the pH was adjusted to 5 with ammonia water. The product was filtered, the filter cake was washed with water, and dried to obtain 14 g of gray-white solid intermediate V-1 with a yield of 94%.

[0130] Synthesis of intermediates V-2 and V-3:

[0131] Compound V-1 (mixture of isomers) (10 g, 27.2 mmol), H3PO3 (9 g, 109.8 mmol), sodium iodide (0.4 g, 2.7 mmol), 50 mL HBr (48%) and 10 mL water were stirred and heated at 105°C for 6 hours, and then cooled to about 100°C. Hypophosphorous acid H3PO2 (50%) (8 mL, 60.6 mmol) was added to the solution, and then heated at 110°C for about 6 hours until the reaction was complete as monitored by HPLC. The reaction solution was cooled to about 90°C, and then acetic acid (20 mL) and ethanol (50 mL) were added. The solution continued to react for 2 hours, and then cooled to 15°C to form a reaction solution containing V2. Liquid bromine (3.3 mL, 63.9 mmol) was dripped into the reaction solution at a temperature of 15 to 20°C, and stirred for another 1 hour. After TLC confirmed that the reaction was complete, post-treatment was performed, and 60 mL 25% ammonia water was slowly added to the mixture to quench the reaction, and then cooled to 25°C and filtered. Wash the filter cake and dry it in water to obtain crude product V-310g, and proceed to the next step without purification.

[0132] Synthesis of intermediate VI:

[0133] Add intermediate V3 (12g, 29.81mmol), 24mL water, and 36mL 98% concentrated sulfuric acid to a three-necked flask, cool to 5-10°C, add hypophosphorous acid (48mL, 16eq), and then add sodium nitrite (2.42g, 1.2eq), heat to 50°C and react for 4h. After the reaction is complete, pour the reaction solution into ice water, neutralize with ammonia water, filter, wash the filter cake with water, and dry. Column chromatography (PE:EA=10:1) gives 5g of white solid intermediate VI.

[0134] Synthesis of intermediate VII:

[0135] 1-Boc-4-methanesulfonyloxypiperidine (3g, 10.74mmol) and intermediate VI (3.5g, 9.05mmol) were dissolved in 15mL toluene, quinine (3.5g, 10.07mmol) was added, the temperature was lowered to 25°C, 13mL lithium diisopropylamide LDA and 0.16mL water were slowly added, and the reaction was complete after 4h of TLC monitoring. Water was added to quench the reaction, the organic layer was separated, the aqueous layer was extracted with ethyl acetate, washed with water, washed with saturated brine, dried and concentrated, and then column chromatography (PE:EA=10:1) was performed to obtain 2g of white solid intermediate VII.

[0136] Synthesis of intermediate VIII:

[0137] 2g of intermediate VII was dissolved in 5 mL of ethyl acetate, 10 mL of hydrogen chloride / ethyl acetate solution was added, and the reaction was carried out at room temperature for 2 hours. After TLC monitoring reaction was completed, the reaction was suction filtered, and after drying, intermediate VIII was obtained as a white solid.

[0138] Synthesis of intermediate IX:

[0139] 2 g of intermediate VIII was dissolved in 5 mL of ethanol, the temperature was raised to reflux for 10 min, and then N-acetyl-L-phenylalanine (2 g, 9.65 mmol) was added. With the addition of N-acetyl-L-phenylalanine, a large amount of white solid precipitated from the reaction solution, and 1.2 g of white solid intermediate IX was obtained by suction filtration.

[0140] Synthesis of intermediate 14-1:

[0141] Intermediate IX (0.5 g, 1.06 mmol) was dissolved in 5 mL of dichloromethane, and chloroacetyl chloride (0.15 g, 1.27 mmol) and TEA (0.22 g, 2.12 mmol) were added under ice bath, followed by reaction at room temperature for 2 h. After the reaction was completed by TLC, saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted three times with DCM. The organic layers were combined, washed with water, and washed once with brine. The concentrated white foamy solid intermediate 14-10.7 g was directly used for the next step without purification.

[0142] Synthesis of compound 14:

[0143] The intermediate 14-1 (0.5 g, 0.91 mmol) was dissolved in 10 mL of acetonitrile, and piperidine-4-carboxamide (0.14 g, 1.09 mmol) and potassium carbonate (0.25 g, 1.93 mmol) were added, followed by reaction at 75 °C for 2 h. After the TLC reaction was complete, water was added, and DCM was extracted three times. The organic layers were combined, washed with water, washed once with brine, concentrated, and purified by column chromatography (DCM:MeOH=50:1) to obtain 0.3 g of white solid compound 14.

[0144] 1 H NMR (300MHz, CDCl3) δ=7.75(d,J=2.2Hz,1H),7.53(d,J=8.6Hz,1H),7.49(dd,J=8.6,2.2Hz,1H),7.40(dd,J= 8.1,1.7Hz,1H),7.35–7.25(m,2H),7.25–7.18(m,1H),5.29(s,2H),3.74–3.67(m,1H),3.70–3.66(m,1H),3.6 6–3.61(m,1H),3.64–3.57(m,3H),3.43(dd,J=5.3,4.0Hz,2H),3.41(s,3H),2.63–2.52(m,4H),2.33(dd,J=1 5.1,6.9Hz,1H),2.25(dd,J=15.2,7.0Hz,1H),2.06–1.96(m,1H),1.88–1.74(m,4H).HR-MS(ESI):Calculated for C 27 H 31 ClN4O4S[M+H] + :637.0575 found 637.0573. Yield 52%.

[0145] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with piperazine-1-formamide hydrochloride (cas: 474711-89-2) to obtain compound 15:

[0146]

[0147] 1H NMR (300MHz, CDCl3) δ=z8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1 H),7.05–6.93(m,1H),5.24(s,2H),3.95–3.84(m,3H),3.47(dd,J=5.0,4.3Hz,2H),3.43 (dd,J=4.9,4.3Hz,2H),3.32–3.20(m,2H),3.17(d,J=15.2Hz,1H),3.07–2.90(m,4H),2. 64(dd,J=5.1,4.2Hz,4H),2.27–2.19(m,1H),1.82–1.64(m,4H).HR-MS(ESI):Calculated for C 26 H 30 Br2ClN5O2[M+H] + :638.0528 found 638.0523.

[0148] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with 3-aminopyrrolidine (cas: 79286-79-6) to obtain compound 18:

[0149]

[0150] 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H) ,7.05–6.94(m,1H),3.95–3.82(m,3H),3.33–3.20(m,3H),3.12(d,J=4.8Hz,2H),3.07–2. 85(m,5H),2.81–2.72(m,2H),2.62(dd,J=12.5,2.2Hz,1H),2.38–2.33(m,1H),2.22(dd,J =7.0,5.3Hz,1H),2.09(dd,J=7.1,5.2Hz,1H),1.84–1.57(m,6H).HR-MS(ESI):Calculated for C 25 H 29 Br2ClN4O[M+H] + :595.0469 found 595.0467.

[0151] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with 3-hydroxyazetidine (cas: 45347-82-8) to obtain compound 19:

[0152]

[0153] 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.05–6.95(m,,1H),3.98–3.82(m,4H),3.28–6 .21(m,2H),3.23–3.17(m,2H),3.11(d,J=12.8Hz,1H),3.07–2.89(m,9H),2.38–2.33(m,1H),1.75–1.69(m,,1H).HR-MS(ESI):Calculated for C 24 H 26 Br2ClN3O2[M+H] + :582.0153 found 582.0150.

[0154] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with 3,6-diazabicyclo[3.1.1]heptane-3-formamide to obtain compound 43:

[0155]

[0156] 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.52 (d, J = 14.9Hz, 2H), 7.14 (d, J = 2.6Hz, 1H), 4.90 (d, J = 10.3Hz, 1H), 4.66(s,2H),4.50(d,J=12.0Hz,1H),3.83(s,3H),3.74–3.53(m,3H),3.30(t,J=4.4Hz,2H),3.24(d,J=4.8 Hz,1H),3.07–2.92(m,1H),2.85(s,1H),2.80(s,1H),2.67(d,J=7.2Hz,1H),2.39(d,J=13.4Hz,2H),2.04( s,2H),1.57–1.53(m,1H),1.51(s,1H),1.44–1.34(m,2H),1.27(d,J=8.4Hz,2H).HR-MS(ESI):Calculated for C 27H 30 Br2ClN5O2[M+H] + :650.0528 found 650.0525.

[0157] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with (1R, 5S)-3,8-diazabicyclo[3.2.1]octane-8-formamide to obtain compound 44:

[0158]

[0159] 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 2.3Hz, 1H), 7.53 (d, J = 14.0Hz, 2H), 7.15 (t, J = 2.6Hz, 1H), 4. 91(d,J=10.3Hz,1H),4.54(s,3H),4.12(s,3H),3.60(d,J=15.0Hz,1H),3.35–3.12(m,2H),3.11 –2.91(m,2H),2.89–2.76(m,2H),2.64(d,J=24.7Hz,2H),2.53–2.29(m,4H),1.92(s,4H),1.77 (s,1H),1.57(s,1H),1.45(s,1H),1.39(s,1H),1.27(d,J=9.2Hz,1H).HR-MS(ESI):Calculated for C 28 H 32 Br2ClN5O2[M+H] + :664.0684found664.0680.

[0160] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with (1R, 5S)-3,8-diazabicyclo[3.2.1]octane-3-formamide to obtain compound 45:

[0161]

[0162] 1H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.63–7.38 (m, 2H), 7.15 (s, 1H), 4.89 (d ,J=10.3Hz,1H),4.56(s,3H),4.11(s,1H),3.72–3.42(m,3H),3.31(s,2H), 3.25(s,4H),3.01(d,J=14.1Hz,1H),2.93–2.80(m,2H),2.43(d,J=14.3Hz,2H),1.99(s,3H),1.61–1.36(m,4H),1.29(s,2H).HR-MS(ESI):Calculated for C 28 H 32 Br2ClN5O2[M+H] + :664.0684found664.0680.

[0163] Using the synthesis method of compound 14 in Example 3, piperidine-4-formamide was replaced with 2,5-diazabicyclo[2.2.1]heptane-2-formamide to obtain compound 47:

[0164]

[0165] 1 H NMR(300MHz, CDCl3)δ=8.44(s,1H),7.55(s,1H),7.50(s,1H),7.15(s,1H),4.95–4.84(m, 1H),4.49(s,3H),4.05–3.89(m,1H),3.74–3.59(m,2H),3.57(s,1H),3.53–3.37(m,2H),3. 28(d,J=19.5Hz,2H),3.01(d,J=16.1Hz,2H),2.89–2.76(m,2H),2.41(d,J=13.9Hz,2H),1. 95(s,1H),1.76(s,1H),1.56–1.32(m,4H),1.27(d,J=9.1Hz,2H).HR-MS(ESI):Calculated for C 27 H 30 Br2ClN5O2[M+H] + :650.0528 found 650.0525.

[0166] Example 4: Synthesis of 4-(2-(4-(R)-3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cyclohexano[1,2-b]pyridin-11-yl)piperidin-1-yl)-1-fluoro-2-oxoethyl)piperidin-1-formamide (23)

[0167]

[0168] Synthesis of 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid (Intermediate 23-2):

[0169]

[0170] Synthesis of intermediate 23-1:

[0171] The raw material tert-butyl 4-(2-ethoxy-2-oxoethyl)piperidine-1-carboxylate (25 g, 92.1 mmol, cas: 135716-09-5) was dissolved in 250 mL of anhydrous THF, replaced with N2 three times, cooled to -78°C, then LDA (55 mL, 110.6 mmol) was added dropwise, stirred for 30 min, and then NFSI (43 g, 138.2 mmol) in THF (100 mL) was slowly added dropwise. After stirring for 1 h, the mixture was reacted at room temperature for 2 h. After TLC confirmed that the reaction was complete, it was filtered, extracted with EA twice, the organic layers were combined, washed with water, washed once with brine, concentrated, then slurried with DCM, the filter cake was discarded, the filtrate was concentrated and purified by column chromatography (PE:EA=15:1) to obtain intermediate 23-110 g with a yield of 35.7%.

[0172] Synthesis of 23-2:

[0173] The intermediate 23-1 was dissolved in ethanol (100 mL), and then 3N NaOH aqueous solution (30 mL) was added and stirred at room temperature for 3 h. After TLC confirmed that the reaction was complete, the pH was adjusted to 4, and the filtrate was extracted with EA three times. The organic layers were combined, washed with water, washed once with brine, and concentrated. Then, the mixture was slurried with isopropyl ether and filtered. The filter cake was slurried with (PE:EA=10:1) and filtered to obtain a white solid intermediate 23-25 ​​g, with a yield of 55.4%. HR-MS (ESI): Calculated for C 12 H 20 FNO4[M-H] － :260.1304 found 260.1294.

[0174] Synthesis of intermediate 23-3:

[0175] The intermediate 23-2 (0.50 g, 1.92 mmol) was dissolved in DMF (10 mL), EDCI (0.73 g, 3.83 mmol), HOBT (0.52 g, 3.83 mmol), and TEA (0.58 g, 5.74 mmol), and stirred at room temperature for 30 min. Then, the intermediate IX (0.75 g, 1.59 mmol) was added and reacted at room temperature overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water, and solid precipitated. The solid was then filtered off with suction. After the filter cake was dried, it was purified by silica gel column chromatography (PE:EA=5:1) to obtain 0.8 g of the intermediate 23-3 with a yield of 58.6%.

[0176] Synthesis of intermediate 23-4:

[0177] The intermediate 23-3 was dissolved in EA (8 mL), placed in an ice bath, and then 4N HCl-EA (4 mL) solution was added. The reaction was allowed to react at room temperature for 1 h. After the reaction was completed as monitored by TLC, it was filtered with suction. The filter cake was freed with 25% aqueous ammonia (4 mL) for 10 min and extracted with EA three times. The organic layers were combined, washed with water, washed once with brine, and concentrated to obtain 0.6 g of white solid intermediate 23-4 with a yield of 87.2%.

[0178] Synthesis of compound 23:

[0179] The intermediate 23-4 (0.1 mmol) was added to a single-necked bottle, and a mixed solvent of NMP:H2O (4:1, 15 mL) was added, followed by urea (0.15 mmol), and the mixture was refluxed at 130°C for 9 h. After the TLC reaction was complete, the temperature was lowered to 25°C, and the reaction solution was poured into water, where a solid precipitated. The solid was then filtered and purified by filter cake column chromatography (DCM:MeOH=50:1) to obtain a white solid compound 23. 1H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.3Hz, 1H), 7.56 (s, 1H), 7.51 (d, J = 2.2Hz, 1H), 7.16 (d, J = 2.3Hz, 1H), 4.92 (d ,J=10.4Hz,1H),4.60–4.49(m,2H),4.02–3.92(m,2H),3.62(t,J=14.2Hz,1H),3.27(dt,J=17.9,4.3Hz,1H),3. 08–2.91(m,2H),2.89–2.83(m,2H),2.83–2.76(m,1H),2.49(s,1H),2.40(d,J=7.2Hz,1H),2.13(s,1H),1.91(s ,2H),1.63(s,2H),1.55(s,1H),1.43(s,2H),1.41–1.33(m,2H),1.27(d,J=8.1Hz,1H).HR-MS(ESI):Calculated for C 27 H 30 Br2ClFN4O2[M+H] + :655.0481 found 655.0483.

[0180] Compound 23 was prepared by chiral preparation of the liquid phase to obtain compounds 25 and 26.

[0181]

[0182] 1 H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.3Hz, 1H), 7.56 (s, 1H), 7.51 (d, J = 2.2Hz, 1H), 7.16 (d, J = 2.3Hz, 1H), 4.92 (d,J=10.4Hz,1H),4.60–4.49(m,2H),4.02–3.92(m,2H),3.62(t,J=14.2Hz,1H),3.27–3.24(m,1H),3.08–2. 91(m,2H),2.89–2.83(m,2H),2.83–2.76(m,1H),2.49(s,1H),2.40(d,J=7.2Hz,1H),2.13(s,1H),1.91(s,2H ),1.63(s,2H),1.55(s,1H),1.43(s,2H),1.41–1.33(m,2H),1.27(d,J=8.1Hz,1H).HR-MS(ESI):Calculated for C 27 H 30Br2ClFN4O2[M+H] + :655.0481 found 655.0475.

[0183]

[0184] 1 H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.3Hz, 1H), 7.56 (s, 1H), 7.51 (d, J = 2.2Hz, 1H), 7.16 (d, J = 2.3Hz, 1H), 4.92 (d,J=10.4Hz,1H),4.60–4.49(m,2H),4.02–3.92(m,2H),3.62(t,J=14.2Hz,1H),3.27–3.24(m,1H),3.08–2. 91(m,2H),2.89–2.83(m,2H),2.83–2.76(m,1H),2.49(s,1H),2.40(d,J=7.2Hz,1H),2.13(s,1H),1.91(s,2H ),1.63(s,2H),1.55(s,1H),1.43(s,2H),1.41–1.33(m,2H),1.27(d,J=8.1Hz,1H).HR-MS(ESI):Calculated for C 27 H 30 Br2ClFN4O2[M+H] + :655.0481 found 655.0477.

[0185] The synthesis method of compound 23 in Example 4 was used to replace 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid with 1-tert-butoxycarbonyl-4-(methoxycarbonylmethyl)piperidine (cas: 175213-46-4) to obtain compound 16:

[0186]

[0187] 1H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6Hz, 1H), 7.78 (d, J = 1.6Hz, 1H), 7.47 (d, J = 2.2Hz, 1H), 7. 00(s,1H),5.28(s,1H),3.92(d,J=9.5Hz,1H),3.72–3.65(m,2H),3.68–3.58(m,2H),3.49–3.4 5(m,1H),3.39–3.28(m,3H),3.07–2.96(m,1H),2.99–2.93(m,1H),2.96–2.89(m,2H),2.43–2 .23(m,4H),1.95–1.75(m,4H),1.78–1.72(m,1H),1.75–1.67(m,1H).HR-MS(ESI):Calculated for C 26 H 29 Br2ClN4O2[M+H] + :623.0419 found 623.0409.

[0188] Using the synthesis method of compound 23 in Example 4, urea was replaced with deuterated urea (cas: 11433-11-0) to obtain compound 17:

[0189]

[0190] 1 H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.78–7.75(d,J=1.6Hz,1H),7.47(d,J =2.2Hz,1H),7.00(m,1H),3.92(d,J=9.5Hz,1H),3.72–3.69(m,2H),3.64–3.61(m,2 H),3.48–3.44(m,2H),3.32(m,2H),3.08–2.89(m,4H),2.43–2.28(m,2H),2.25(dd, J=15.2,7.0Hz,1H),2.06–1.96(m,1H),1.89–1.67(m,8H).HR-MS(ESI):Calculated forC 27 H 29 D2Br2ClN4O2[M+H] + :639.0701found639.0677.

[0191] Using the synthesis method of compound 23 in Example 4, urea was replaced with (E)-2-(hydroxyimino)acetic acid to obtain compound 20:

[0192]

[0193] 1 H NMR (300MHz, CDCl3) δ = 8.47 (s, 1H), 7.77 (s, 1H), 7.62 (d, J = 13.5Hz, 1H), 7.51 (d, J = 2.2Hz, 1 H),7.15(s,1H),4.94(d,J=10.0Hz,1H),4.61(d,J=13.1Hz,2H),4.23(s,1H),3.84(s,1H),3 .63(s,1H),3.29(d,J=17.1Hz,1H),3.06(s,2H),2.84(d,J=14.8Hz,2H),2.72(s,1H),2.24( d,J=7.2Hz,2H),1.83(s,3H),1.37(s,2H),1.23(d,J=15.8Hz,3H).HR-MS(ESI):Calculated for C 28 H 31 Br2ClN4O3[M+H] + :665.0524 found 665.0520.

[0194] Using the synthesis method of compound 23 in Example 4, urea was replaced with (E)-2-(methoxyimino)acetic acid to obtain compound 21:

[0195]

[0196] 1 H NMR (300MHz, CDCl3) δ = 8.51 (s, 1H), 7.70 (t, J = 11.5Hz, 2H), 7.52 (d, J = 2.2Hz, 1H), 7.17 (s, 1H), 5. 01(d,J=9.6Hz,1H),4.63(s,2H),4.19(s,1H),4.03–3.93(m,3H),3.85(s,1H),3.64(s,1H),3.32( d,J=17.3Hz,1H),3.21–2.98(m,2H),2.85(d,J=13.0Hz,2H),2.73(s,1H),2.41(s,2H),2.19(d,J= 23.4Hz,3H),1.85(t,J=13.1Hz,2H),1.41(d,J=18.6Hz,3H),1.26(s,3H).HR-MS(ESI):Calculated for C 29 H 33 Br2ClN4O3[M+H]+ :679.0681 found 679.0671.

[0197] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2,2-difluoroacetic acid to obtain compound 24:

[0198]

[0199]

[0200] 1 H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00–6.97(m,1H),5.29(s,1H),3.95–3.73(m,4H),3.62 –3.53(m,3H),3.07–2.96(m,1H),2.99–2.94(m,1H),2.96–2.90(m,1H),2.74–2.71(m,1H),2.21(m,1H),1.89–1.69(m,7H).HR-MS(ESI):Calculated for C 27 H 29 Br2ClF2N4O2[M+H] + :673.0387found673.0380.

[0201] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 4-carbamoylbenzoic acid to obtain compound 27:

[0202]

[0203] 1H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),8.01–7.95(m,2H),7.78(d,J=1.6Hz,1H),7.53–7.45(m,3H),7.00(s,1H),6.90(s,1H),3.9 9–3.90(m,3H),3.34–3.30(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.20–2.18(m,1H),1.86–1.70(m,4H).HR-MS(ESI):Calculated for C 27 H 24 Br2ClN3O2[M+H] + :614.9924 found 614.9920.

[0204] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 4-carbamoyl-2-fluorobenzoic acid to obtain compound 28:

[0205]

[0206] 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6 Hz, 1H), 7.96 (dd, J = 8.3, 1.9 Hz, 1H), 7.86 ( dd,J=7.9,1.9Hz,1H),7.78(d,J=1.6Hz,1H),7.52–7.44(m,3H),7.42(d,J=8.1H z,1H),7.00(s,1H),3.99–3.90(m,3H),3.81–3.78(m,2H),3.07–2.94(m,2H),2. 98–2.90(m,2H),2.20–2.18(m,1H),1.87–1.70(m,4H).HR-MS(ESI):Calculated for C 27 H 23 Br2ClFN3O2[M+H] + :633.9902 found 633.9912.

[0207] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 4-carbamoyl-2-methoxybenzoic acid to obtain compound 29:

[0208]

[0209] 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6Hz, 1H), 7.92 (dd, J = 8.6, 1.8Hz, 1H), 7.78 (d, J=1.6Hz,1H),7.57(d,J=1.9Hz,1H),7.52–7.37(m,4H),7.00(s,1H),3.99–3.92(m ,3H),3.95–3.91(m,1H),3.94–3.89(m,1H),3.81–3.78(m,2H),3.07–2.98(m,1H), 3.00–2.90(m,3H),2.20–2.18(m,1H),1.87–1.70(m,4H).HR-MS(ESI):Calculated forC 28 H 26 Br2ClN3O3[M+H] + :646.0102found646.0100.

[0210] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 2-(4-carbamoylphenyl)acetic acid to obtain Compound 31:

[0211]

[0212] 1 H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.92–7.86(m,2H),7.78(d,J=1.6Hz ,1H),7.47(d,J=2.2Hz,1H),7.29(dt,J=8.6,1.0Hz,2H),7.00(s,1H),6.91(s,2H) ,3.95–3.84(m,3H),3.66(t,J=1.0Hz,2H),3.25–3.21(m,2H),3.07–2.98(m,1H), 3.00–2.90(m,3H),2.21–2.18(m,1H),1.78–1.68(m,4H).HR-MS(ESI):Calculated forC 28 H 26 Br2ClN3O2[M+H] + :630.0153found630.0150.

[0213] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 2-(4-carbamoylphenyl)-2,2-difluoroacetic acid to obtain compound 32.

[0214]

[0215]

[0216] 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.94–7.88(m,2H),7.78(d,J=1.6Hz,1H),7.57–7.50(m,2H),7.47(d,J=2.2Hz,1H),7.00(s,1H),6.91 (s,1H),3.95–3.85(m,3H),3.53–3.49(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.21–2.18(m,1H),1.84–1.68(m,4H).HR-MS(ESI):Calculated for C 28 H 24 Br2ClF2N3O2[M+H] + :665.9965found665.9960. .

[0217] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 2-(4-carbamoylphenyl)-2,2-dimethylacetic acid to obtain compound 33.

[0218]

[0219] 1 H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.83–7.76(m,3H),7.47(d,J=2.2Hz,1H),7.32–7.26(m,2H),7.00(s,1H),6. 91(s,1H),3.92(d,J=9.5Hz,1H),3.84–3.81(m,2H),3.29–3.25(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.21–2.18(m 1H),1.83–1.66(m,4H).HR-MS(ESI):Calculated for C 30 H 30Br2ClN3O2[M+H] + :658.0466found658.0461.

[0220] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 1-(4-carbamoylphenyl)cyclopropane-1-carboxylic acid to obtain compound 34.

[0221]

[0222] 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6 Hz, 1H), 7.81–7.75 (m, 3H), 7.47 (d, J = 2. 2Hz,1H),7.38–7.32(m,2H),7.00(s,1H),6.91(s,2H),3.92(d,J=9.5Hz,1H),3 .86–3.83(m,2H),3.27–3.23(m,2H),3.07–2.90(m,4H),2.21–2.18(m,J=9.4,7 .1,4.6Hz,1H),1.83–1.60(m,7H),1.50–1.40(m,2H).HR-MS(ESI):Calculated for C 30 H 28 Br2ClN3O2[M+H] + :656.0310 found 656.0305.

[0223] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 2-(4-carbamoyl-3-fluorophenyl)acetic acid to obtain compound 35.

[0224]

[0225] 1H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6Hz, 1H), 7.82 (dd, J = 8.5, 5.0Hz, 1H), 7.78 (d, J = 1. 7Hz,1H),7.57(d,J=7.9Hz,1H),7.49–7.41(m,2H),7.35–7.32(m,1H),7.28–7.25(m,1H) ,7.00(s,1H),3.95–3.84(m,3H),3.66(t,J=1.0Hz,2H),3.25–3.21(m,2H),3.07–2.94(m ,2H),2.98–2.90(m,2H),2.21–2.18(m,1H),1.78–1.68(m,4H).HR-MS(ESI):Calculated for C 28 H 25 Br2ClFN3O2[M+H] + :648.0059found 648.0055.

[0226] Using the synthesis method of compound 23 in Example 4, 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid was replaced with 2-(4-(hydroxycarbamoyl)phenyl)acetic acid to obtain compound 36.

[0227]

[0228] 1 H NMR (300MHz, CDCl3) δ = 9.94 (d, J = 3.5Hz, 1H), 8.66 (d, J = 3.5Hz, 1H), 8.44 (d, J = 1.6Hz, 1H),7.85–7.80(m,2H),7.78(d,J=1.7Hz,1H),7.47(d,J=2.2Hz,1H),7.37(s,2H),7.00 (s,1H),3.95–3.84(m,3H),3.66(t,J=1.0Hz,2H),3.25–3.21(m,2H),3.07–2.96(m,1H) ,2.99–2.90(m,3H),2.21–2.18(m,1H),1.78–1.68(m,4H).HR-MS(ESI):Calculatedfor C 28 H 26 Br2ClN3O3[M+H] + :646.0102 found 646.0100.

[0229] The synthesis method of compound 23 in Example 4 was used to replace 2-(2-(tert-butoxycarbonyl)-2-aza[3.3]heptane-6-yl)acetic acid with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid to obtain compound 37 (cas: 1251002-39-7).

[0230]

[0231] 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s ,1H),5.03(d,J=7.0Hz,1H),4.96(d,J=6.8Hz,1H),3.92(d,J=9.5Hz,1H),3.76(s,2H),3.61–3.57( m,2H),3.57(s,2H),3.31–3.28(m,2H),3.07–2.90(m,4H),2.33–2.31(m,1H),2.29–2.16(m,2H),2. 04–1.92(m,1H),1.82–1.74(m,1H),1.77–1.65(m,5H),1.56–1.51(m,2H).HR-MS(ESI):Calculated for C 28 H 31 Br2ClN4O2[M+H] + :649.0575 found 649.0570.

[0232] The synthesis method of compound 23 in Example 4 was adopted to replace 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid with 1-carbamoylpiperidine-4-carboxylic acid to obtain compound 41.

[0233]

[0234] 1H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s,1H),5.29(s,1H),3.92(d,J=9.5Hz,1H),3.73–3.5 8(m,4H),3.37–3.27(m,4H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.55–2.51(m,1H),2.21–2.18(m,1H),1.87–1.65(m,8H).HR-MS(ESI):Calculated for C 26 H 29 Br2ClN4O2[M+H] + :623.0419found623.0410.

[0235] The synthesis method of compound 23 in Example 4 was adopted to replace 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid with thymine-1-acetic acid (cas: 20924-05-4) to obtain compound 42.

[0236]

[0237] 1 H NMR (300MHz, CDCl3) δ = 9.83 (s, 1H), 8.44 (d, J = 1.6Hz, 1H), 7.78 (d, J = 1.6Hz, 1H),7.47(d,J=2.2Hz,1H),7.26(d,J=1.5Hz,1H),7.00(s,1H),4.50(s,1H), 3.95–3.85(m,3H),3.37–3.32(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H), 2.21–2.18(m,1H),1.87(s,1H),1.82–1.64(m,4H).HR-MS(ESI):Calculated for C 26 H 25 Br2ClN4O3[M+H] + :635.0055found 635.0050.

[0238] The synthesis method of compound 23 in Example 4 was adopted to replace 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid with 2-(4-sulfonylphenyl)acetic acid to obtain compound 46.

[0239]

[0240] 1 H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.83–7.76(m,3H),7.47(d,J=2.2Hz,1H),7.40–7.35(m,2H),7.06(s,2H),7.00(s,1H),3.95–3.84(m, 3H),3.65(t,J=1.0Hz,2H),3.25–3.21(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.21–2.18(m,1H),1.78–1.68(m,4H).HR-MS(ESI):Calculated for C 27 H 26 Br2ClN3O3S[M+H] + :665.9823found665.9820.

[0241] The synthesis method of compound 23 in Example 4 was adopted to obtain compound 49 by replacing 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid with 2-(5-carbamoylpyrimidin-2-yl)acetic acid.

[0242]

[0243] 1 H NMR (300MHz, CDCl3) δ = 9.59 (s, 1H), 8.44 (d, J = 1.6Hz, 1H), 8.11 (d, J = 8.1Hz, 1H), 7.82–7.76 (m, 2H), 7.47 (d, J = 2.2Hz, 1H), 7.00 (s, 1H), 3.95–3. 85(m,3H),3.55(s,1H),3.27–3.23(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.21–2.18(m,1H),1.82–1.65(m,4H).HR-MS(ESI):Calculated for C 26 H 24 Br2ClN5O2[M+H] + :632.0058 found 632.0054.

[0244] The synthesis method of compound 23 in Example 4 was adopted to replace 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid with 1-N-Boc-4-piperidinepropionic acid (cas: 154775-43-6) to obtain compound 52.

[0245]

[0246] 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6 Hz, 1H), 7.78 (d, J = 1.6 Hz, 1H), 7.47 (d, J = 2.2 Hz, 1H),7.00(s,1H),5.29(s,2H),3.92(d,J=9.5Hz,1H),3.80–3.75(m,2H),3.71–3.61(m ,2H),3.60–3.51(m,2H),3.31–3.26(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.2 8(t,J=8.8Hz,2H),2.21–2.18(m,1H),1.82–1.51(m,12H).HR-MS(ESI):Calculatedfor C 28 H 33 Br2ClN4O2[M+H] + :651.0732 found 651.0722.

[0247] Example 5: Synthesis of (R)-2-(4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidin-1-yl)acetamide (48)

[0248]

[0249] Synthesis of intermediate 48-1:

[0250] 1-tert-Butyloxycarbonyl-4-piperidinylacetic acid (0.50 g, 1.92 mmol, cas: 157688-46-5), EDCI (0.73 g, 3.83 mmol), HOBT (0.52 g, 3.83 mmol), TEA (0.58 g, 5.74 mmol) were dissolved in DMF (10 mL), stirred at room temperature for 30 min, and then intermediate IX (0.75 g, 1.59 mmol) was added. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water. Solids precipitated and then filtered. After the filter cake was dried, it was purified by silica gel column chromatography (PE:EA=5:1) to obtain intermediate 48-10.8 g with a yield of 59.6%.

[0251] Synthesis of intermediate 48-2:

[0252] The intermediate 23-3 was dissolved in EA (8 mL), placed in an ice bath, and then 4N HCl-EA (4 mL) solution was added. The reaction was allowed to react at room temperature for 1 h. After the reaction was completed as monitored by TLC, it was filtered with suction. The filter cake was freed with 25% aqueous ammonia (4 mL) for 10 min and extracted with EA three times. The organic layers were combined, washed with water, washed once with brine, and concentrated to obtain a white solid intermediate 48-20.6 g with a yield of 87.2%.

[0253] Synthesis of compound 48:

[0254] The intermediate 48-2 (150 mg, 0.23 mmol) was added to a single-mouth bottle, followed by acetonitrile (5 mL), followed by 2-bromoacetamide (38.9 mg, 0.28 mmol, cas: 683-57-8) and potassium carbonate (64.8 mg, 0.46 mmol), and the mixture was reacted at 80 °C for 2 h. After the TLC reaction was complete, the mixture was cooled to room temperature, water was added, and EA was extracted 3 times. The organic layers were combined, washed with water, washed once with brine, concentrated, and purified by column chromatography to obtain white solid compound 48. 1 H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.4Hz, 1H), 7.59–7.53 (m, 1H), 7.50 (d, J = 2.3Hz, 1H), 7.15 (t, J = 2.6Hz, 2H) ,5.54(s,1H),4.90(dd,J=10.5,2.8Hz,1H),4.59(d,J=12.5Hz,1H),3.85(d,J=13.0Hz,1H),3.62(t,J=13.5Hz ,1H),3.30–3.23(m,1H),3.01(s,2H),2.93–2.80(m,4H),2.39(q,J=8.9,4.7Hz,2H),2.23(td,J=11.6,9.6,6. 1Hz,4H),1.50(ddd,J=24.1,14.1,8.0Hz,4H),1.36–1.31(m,2H),1.30–1.22(m,4H).HR-MS(ESI):Calculated for C 28 H 33 Br2ClN4O2[M+H] + :651.0732found651.0722.

[0255] The synthesis method of compound 48 in Example 5 was adopted to obtain compound 53 by replacing 2-bromoacetamide with 3-bromopropylamine hydrobromide (cas: 5003-71-4).

[0256]

[0257] 1H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s,1 H),3.92(d,J=9.5Hz,1H),3.61–6.57(m,2H),3.31–3.27(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H ),2.84–2.78(m,1H),2.81–2.72(m,3H),2.52–2.48(m,2H),2.47(t,J=5.7Hz,2H),2.31(s,1H),2.27 –2.16(m,2H),1.95–1.92(m,1H),1.82–1.62(m,11H),1.53(t,J=6.5Hz,2H).HR-MS(ESI):Calculated for C 29 H 37 Br2ClN4O[M+H] + :651.1095found651.1090.

[0258] The synthesis method of compound 48 in Example 5 was adopted to obtain compound 54 by replacing 2-bromoacetamide with 3-bromopropanol (cas: 627-18-9).

[0259]

[0260] 1 H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s,1H),3 .92(d,J=9.5Hz,1H),3.78–3.72(m,2H),3.61–3.57(m,2H),3.40(t,J=5.9Hz,1H),3.31–3.26(m,2H),3.07 –2.98(m,1H),3.00–2.90(m,3H),2.80–2.75(m,2H),2.60–2.52(m,3H),2.55–2.49(m,1H),2.31–2.28(m, 1H),2.27–2.16(m,2H),1.95–1.92(m,1H),1.82–1.65(m,9H),1.61–1.56(m,2H).HR-MS(ESI):Calculated forC 29 H 36 Br2ClN3O2[M+H] +:652.0936 found 652.0930.

[0261] The synthesis method of compound 48 in Example 5 was adopted to obtain compound 55 by replacing 2-bromoacetamide with 1-bromo-3-(methylsulfonyl)propane (cas: 859940-73-1).

[0262]

[0263] 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.0 0(s,1H),3.92(d,J=9.5Hz,1H),3.61–3.56(m,2H),3.31–3.27(m,2H),3.10(t,J=10.5Hz,2H), 3.07–2.99(m,1H),3.00(s,3H),2.98–2.90(m,3H),2.79–2.72(m,2H),2.56(d,J=7.1Hz,2H),2 .56–2.49(m,2H),2.31(s,1H),2.27–2.16(m,2H),2.00–1.65(m,12H).HR-MS(ESI):Calculated for C 30 H 38 Br2ClN3O3S[M+H] + :714.0762 found 714.0760.

[0264] The synthesis method of compound 48 in Example 5 was adopted to obtain compound 56 by replacing 2-bromoacetamide with 4-bromo-1,1,1-trifluorobutane (cas: 406-81-5).

[0265]

[0266] 1H NMR(300MHz, CDCl3)δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7 .00(s,1H),3.92(d,J=9.5Hz,1H),3.61–3.57(m,2H),3.31–3.27(m,5.9Hz,2H),3.07–2.98(m ,1H),3.00–2.90(m,3H),2.79–2.72(m,2H),2.57–2.49(m,2H),2.53–2.48(m,2H),2.39–2.2 6(m,2H),2.28–2.16(m,3H),2.00–1.89(m,1H),1.82–1.56(m,11H).HR-MS(ESI):Calculated for C 30 H 35 Br2ClF3N3O[M+H] + :704.0860 found 704.0865.

[0267] Example 6: Synthesis of 4-(2-((1R,5S)-3-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)-8-azabicyclo[3.2.1]octan-8-yl)-2-oxoethyl)piperidine-1-carboxamide (30)

[0268]

[0269] Synthesis of intermediate 30-1:

[0270] The raw material tert-butyl 3-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylate (1.0 g, 4.4 mmol, cas: 478837-18-2) was dissolved in DCM (10 mL), cooled to 0°C in an ice bath, and then methanesulfonic anhydride (1.15 g, 6.6 mmol) was added, and TEA (1.22 mL, 8.8 mmol) was slowly added, and then reacted at room temperature for 30 min. After TLC confirmed that the reaction was complete, saturated sodium bicarbonate aqueous solution was added, and DCM was extracted twice, the organic layers were combined, washed with water, washed once with brine, and concentrated to obtain a yellow solid, and then slurried with n-heptane to obtain a white solid intermediate 30-1. The yield was 90%. 1H NMR (300MHz, CDCl3) δ = 5.03 (t, J = 5.0Hz, 1H), 4.22 (s, 2H), 3.01 (s, 3H), 2.21 (dt, J = 15.6, 4.4Hz, 2H), 2.13–1.89 (m, 6H), 1.46 (s, 9H). HR-MS (ESI): Calculated for C 13 H 23 NO5S[M+Na] + :328.1297 found 328.1294.

[0271] Synthesis of intermediate 30-2:

[0272] 30-1 (3.2 g, 10.74 mmol) and intermediate VI (3.5 g, 9.05 mmol) were dissolved in 15 mL of toluene, cooled to 25 °C, and 13 mL of lithium diisopropylamide LDA and 0.16 mL of water were slowly added. After 4 h, the reaction was complete as monitored by TLC. Water was added to quench the reaction, and the organic layer was separated. The aqueous layer was extracted with ethyl acetate, washed with water and saturated brine, dried, concentrated, and then column chromatography (PE:EA=10:1) was performed to obtain 1.5 g of white solid intermediate VII.

[0273] Synthesis of intermediate 30-3:

[0274] 1.5 g of intermediate VII was dissolved in 15 mL of ethyl acetate, and 10 mL of hydrogen chloride / ethyl acetate solution was added. The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the mixture was filtered and dried to obtain a white solid intermediate 30-3.

[0275] Synthesis of intermediate 30-4:

[0276] 1-tert-Butyloxycarbonyl-4-piperidinylacetic acid (0.50 g, 1.92 mmol, cas: 157688-46-5), EDCI (0.73 g, 3.83 mmol), HOBT (0.52 g, 3.83 mmol), TEA (0.58 g, 5.74 mmol) were dissolved in DMF (10 mL), stirred at room temperature for 30 min, and then intermediate 30-3 (0.8 g, 1.59 mmol) was added. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water, and a solid precipitated. The solid was then filtered off with suction. After the filter cake was dried, it was purified by silica gel column chromatography (PE:EA=5:1) to obtain intermediate 30-40.76 g.

[0277] Synthesis of intermediate 30-5:

[0278] The intermediate 30-3 was dissolved in EA (8 mL), placed in an ice bath, and then 4N HCl-EA (4 mL) solution was added. The reaction was allowed to react at room temperature for 1 h. After the reaction was completed as monitored by TLC, the mixture was filtered off with suction. The filter cake was freed with 25% aqueous ammonia (4 mL) for 10 min and extracted with EA three times. The organic layers were combined, washed with water, washed once with brine, and concentrated to obtain 0.6 g of white solid intermediate 30-5 with a yield of 87.2%.

[0279] Synthesis of compound 30:

[0280] The intermediate 23-4 (0.1 mmol) was added to a single-necked bottle, and a mixed solvent of NMP:H2O (4:1, 15 mL) was added, followed by urea (0.15 mmol), and the mixture was refluxed at 130°C for 9 h. After the TLC reaction was complete, the temperature was lowered to 25°C, and the reaction solution was poured into water, where a solid precipitated. The solid was then filtered and purified by filter cake column chromatography (DCM:MeOH=50:1) to obtain a white solid compound 30. 1 H NMR (300MHz, CDCl3) δ = 8.43 (s, 1H), 7.55 (d, J = 9.7Hz, 1H), 7.48 (d, J = 3.5Hz, 1H), 7.13 (s, 1H),4.87(d,J=9.9Hz,1H),4.66(d,J=12.6Hz,2H),4.12(s,1H),3.95(s,2H),3.68–3.52(m ,1H),3.27(d,J=17.4Hz,1H),3.00(s,2H),2.90–2.67(m,4H),2.27–2.05(m,4H),1.83(d, J=15.4Hz,6H),1.59(s,2H),1.51(s,1H),1.23(d,J=22.0Hz,4H).HR-MS(ESI):Calculated for C 29 H 33 Br2ClN4O2[M+H] + :663.0659found663.0663.

[0281] The synthesis method of compound 30 in Example 6 was adopted to replace tert-butyl 3-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylate with tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate (cas: 373604-28-5) to obtain compound 22.

[0282]

[0283] 1H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s, 1H),5.29(s,1H),4.73(d,J=10.6Hz,1H),3.77–3.57(m,4H),3.46–3.32(m,1H),3.07–2.95(m,1H),2 .98–2.89(m,2H),2.64–2.59(m,1H),2.29(d,J=6.8Hz,1H),2.15–2.10(m,1H),2.07–1.97(m,1H),1 .92–1.83(m,1H),1.86–1.81(m,1H),1.83–1.79(m,1H),1.81–1.74(m,1H).HR-MS(ESI):Calculated for C 27 H 30 Br2ClFN4O2[M+H] + :655.0481found655.0475.

[0284] Example 7: Synthesis of (R)-4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)-N-methylpiperidine-1-carboxamide (57)

[0285]

[0286] The synthesis of intermediate 48-2 is the same as in Example 5.

[0287] Synthesis of compound 57:

[0288] The intermediate 48-2 (0.1 mmol) was dissolved in DCM (5 mL), and TEA (0.2 mmol) was added dropwise under an ice bath, followed by the addition of methylaminoformyl chloride (0.12 mmol, cas: 463-72-9), followed by reaction at room temperature for 2 h. After TLC confirmed that the reaction was complete, saturated NaHCO3 aqueous solution was added to quench, followed by extraction with DCM twice, the organic layers were combined, washed with water, washed with brine, concentrated, and purified by column chromatography (DCM:MeOH=30:1) to obtain white solid compound 57 with a yield of 90.1%. 1H NMR (300MHz, CDCl3) δ = 8.47 (s, 1H), 7.59 (d, J = 9.6Hz, 1H), 7.51 (d, J = 2.2Hz, 1H), 7.15 (s, 1H), 4.94 (d, J = 10.2Hz, 1H) ,4.61(d,J=14.0Hz,2H),3.85(dd,J=26.3,13.6Hz,3H),3.70–3.52(m,1H),3.29(d,J=17.7Hz,1H),3.03(d,J=14.3Hz ,1H),2.91–2.84(m,2H),2.81(s,3H),2.37(d,J=11.7Hz,2H),2.23(t,J=6.2Hz,2H),2.03(d,J=11.7Hz,1H),1.76(d, J=12.9Hz,2H),1.53(s,2H),1.40(d,J=15.6Hz,2H),1.26(s,1H),1.16(d,J=11.8Hz,2H).HR-MS(ESI):Calculatedfor C 28 H 33 Br2ClN4O2[M+H] + :651.0732found651.0728.

[0289] The synthesis method of compound 57 in Example 7 was adopted to obtain compound 58 by replacing methylaminoformyl chloride with dimethylaminoformyl chloride (cas: 79-44-7).

[0290]

[0291] 1 H NMR (300MHz, CDCl3) δ = 8.48 (s, 1H), 7.61 (d, J = 8.4Hz, 1H), 7.54–7.47 (m, 1H), 7.15 (s, 1H), 4 .96(d,J=9.8Hz,1H),4.60(d,J=12.6Hz,1H),3.85(s,1H),3.65(d,J=13.2Hz,3H),3.29(d,J= 17.5Hz,1H),3.01(s,1H),2.81(s,11H),2.38(s,2H),2.23(d,J=6.4Hz,2H),1.73(d,J=12.6 Hz,2H),1.51(d,J=17.0Hz,2H),1.43(s,2H),1.35(d,J=9.7Hz,2H).HR-MS(ESI):Calculated for C 29 H 35 Br2ClN4O2[M+H]+ :665.0888found665.0880.

[0292] The synthesis method of compound 57 in Example 7 was adopted to replace 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid to obtain compound 63.

[0293]

[0294] 1 H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.4Hz, 1H), 7.57 (d, J = 4.1Hz, 1H), 7.50 (d, J = 2.3Hz, 1H), 7.16 (d, J = 2.3Hz, 1H), 4 .94(s,1H),4.74(d,J=5.9Hz,1H),4.55(s,2H),4.02(s,1H),3.98(s,2H),3.62(t,J=14.0Hz,1H),3.28(dt,J=18.3 ,4.5Hz,1H),3.07–2.93(m,1H),2.92–2.82(m,2H),2.80(s,3H),2.75(d,J=12.2Hz,2H),2.45(d,J=26.9Hz,2H),2. 07(s,2H),1.92–1.80(m,1H),1.57(d,J=12.7Hz,2H),1.40(s,2H),1.33(d,J=11.4Hz,2H).HR-MS(ESI):Calculated for C 28 H 32 Br2ClFN4O2[M+H] + :669.0637found669.0633.

[0295] The synthesis method of compound 57 in Example 7 was adopted to obtain compound 65 by replacing 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-((1R,5S)-8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)acetic acid.

[0296]

[0297] Synthesis of 2-((1R, 5S)-8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)acetic acid:

[0298]

[0299] 3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester (0.5 g, cas: 201162-53-0) was added to a single-mouth bottle, followed by acetonitrile (5 mL), followed by bromoacetic acid (0.05 g, cas: 79-08-3) and potassium carbonate (0.1 g), and the mixture was reacted at 80°C for 2 h. After the TLC reaction was complete, the mixture was cooled to room temperature, water was added, and DCM was extracted three times. The organic layers were combined, washed with water, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain a colorless oil, which was directly used for the next step without purification.

[0300] Synthesis of compound 66:

[0301] 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.59–7.46 (m, 2H), 7.15 (s, 1H), 4.89 (d, J = 10.3Hz, 1H), 4.50 (s, 2H) ,4.12(t,J=13.3Hz,1H),3.65(d,J=12.9Hz,1H),3.56–3.43(m,2H),3.30(s,2H),3.24(s,2H),3.20–3.07 (m,2H),3.01(d,J=14.4Hz,1H),2.92–2.86(m,1H),2.80(d,J=4.3Hz,3H),3.56–3.34(m,2H),2.18(s,2H) ,2.02–1.92(m,2H),1.70(d,J=9.5Hz,2H),1.43(s,2H),1.35(d,J=12.0Hz,2H).HR-MS(ESI):Calculated forC 29 H 34 Br2ClN5O2[M+H] + :678.0841found678.0839.

[0302] The synthesis method of compound 57 in Example 7 was used to replace 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(2-(tert-butoxycarbonyl)-2-azaspiro[3.3]heptane-6-yl)acetic acid (cas: 1251002-39-7) to obtain compound 64.

[0303]

[0304] 1H NMR (300MHz, CDCl3) δ = 8.46 (s, 1H), 7.61 (s, 1H), 7.51 (s, 1H), 7.16 (s, 1H), 5.08 (s, 1H), 4.93 (d, J = 9.2Hz, 1 H),4.54(d,J=13.0Hz,1H),3.80(s,1H),3.67(s,1H),3.50(d,J=2.9Hz,2H),3.43(s,2H),3.27(s,1H),3.00( s,1H),2.84(d,J=13.4Hz,2H),2.77(s,3H),2.63(s,1H),2.44(d,J=6.6Hz,3H),2.36(d,J=13.7Hz,2H),2.0 8(s,1H),2.04(s,1H),1.66(d,J=10.0Hz,2H),1.43(s,2H),1.35(d,J=10.9Hz,2H).HR-MS(ESI):Calculated for C 29 H 33 Br2ClN4O2[M+H] + :663.0732 found 663.0730.

[0305] The synthesis method of compound 57 in Example 7 was adopted to obtain compound 66 by replacing 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(7-(tert-butoxycarbonyl)-4,7-diazaspiro[2.5]octan-4-yl)acetic acid.

[0306]

[0307] Synthesis of 2-(7-(tert-butoxycarbonyl)-4,7-diazaspiro[2.5]octan-4-yl)acetic acid:

[0308]

[0309] Add tert-butyl 4,7-diazaspiro[2.5]octane-7-carboxylate (0.5 g, cas: 886766-28-5) to a single-mouth bottle, then add acetonitrile (5 mL), followed by bromoacetic acid (0.05 g, cas: 79-08-3) and potassium carbonate (0.1 g), and react at 80°C for 2 h. After the TLC reaction is complete, cool to room temperature, add water, extract with DCM three times, combine the organic layers, wash with water, wash once with brine, dry over anhydrous sodium sulfate, and concentrate to obtain a colorless oil, which is directly used for the next step without purification.

[0310] Synthesis of compound 66:

[0311] 1H NMR (300MHz, CDCl3) δ = 8.46 (s, 1H), 7.61 (s, 1H), 7.51 (s, 1H), 7.16 (s, 1H), 5.08 (s, 1H), 3.95–3 .84(m,3H),3.54(s,1H),3.46–3.40(m,1H),3.41(s,1H),3.36–3.30(m,1H),3.32–3.24(m,2H),3 .25(s,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.86–2.80(m,1H),2.82–2.75(m,1H),2.74(d, J=5.5Hz,3H),2.21–2.16(m,1H),1.92–1.84(m,2H),1.87–1.64(m,6H).HR-MS(ESI):Calculated for C 29 H 34 Br2ClN5O2[M+H] + :678.0841found678.0843.

[0312] The synthesis method of compound 57 in Example 7 was used to obtain compound 67 by replacing 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(4-(tert-butoxycarbonyl)piperazine-1-yl)acetic acid (cas: 156478-71-6).

[0313]

[0314] 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.52 (d, J = 14.8Hz, 2H), 7.15 (s, 1H), 4.89 (d, J = 10.2Hz, 1H), 4.52(s,2H),4.02(d,J=12.1Hz,1H),3.62(t,J=14.1Hz,1H),3.40(s,4H),3.26(d,J=16.6Hz,2H), 3.14(t,J=13.9Hz,1H),2.98(t,J=15.5Hz,1H),2.85(s,2H),2.81(d,J=4.4Hz,3H),2.52(s,4H),2 .38(d,J=16.2Hz,2H),2.09(s,1H),1.54(d,J=12.8Hz,2H),1.40(s,2H).HR-MS(ESI):Calculated for C 27 H 32 Br2ClN5O2[M+H] +:652.0684 found 652.0680.

[0315] The synthesis method of compound 57 in Example 7 was used to obtain compound 68 by replacing 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(1-(tert-butoxycarbonyl)-4-hydroxypiperidin-4-yl)acetic acid (cas: 502482-52-2).

[0316]

[0317] 1 H NMR (300MHz, CDCl3) δ = 8.46 (s, 1H), 7.61 (s, 1H), 7.51 (s, 1H), 7.16 (s, 1H), 5.08 (s, 1H) ),4.13(s,1H),3.92(d,J=9.5Hz,1H),3.84–3.76(m,2H),3.53–3.46(m,2H),3.40–3.26 (m,4H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.74(d,J=5.5Hz,3H),2.21–3.17(m,1H) ,2.01–1.95(m,2H),1.85–1.77(m,2H),1.81–1.66(m,4H).HR-MS(ESI):Calculatedfor C 28 H 33 Br2ClN4O3[M+H] + :667.0681found667.0687.

[0318] Example 8: Synthesis of (R)-N-cyclopropyl-4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidine-1-carboxamide (60)

[0319]

[0320] The intermediate 48-2 (100 mg, 0.17 mmol) was dissolved in DCM (5 mL), TEA (44.5 μL) was added dropwise under ice bath, followed by cyclopropyl isocyanate (16.74 mg, 0.20 mmol, cas: 4747-72-2), and then reacted at room temperature for 2 h. After TLC confirmed that the reaction was complete, saturated NaHCO3 aqueous solution was added to quench, and then DCM was extracted twice, the organic layers were combined, washed with water, washed with brine, concentrated, and purified by column chromatography (DCM: MeOH = 40: 1) to obtain a white solid compound 64. The yield was 80%. 1HNMR (300MHz, CDCl3) δ = 8.46 (s, 1H), 7.62–7.48 (m, 2H), 7.16 (s, 1H), 4.93 (d, J = 10.2Hz, 1H), 4.74 (s, 1H), 4.61 ( d,J=13.2Hz,1H),3.87(d,J=15.0Hz,3H),3.61(d,J=14.7Hz,1H),3.28(d,J=17.6Hz,1H),3.02(d,J=15.4Hz,1H), 2.81(t,J=15.2Hz,4H),2.65(s,1H),2.39(s,1H),2.26–2.18(m,2H),2.00(s,1H),1.75(d,J=12.8Hz,2H),1.40(d ,J=15.7Hz,2H),1.16(s,1H),0.89(t,J=6.5Hz,2H),0.72(d,J=6.6Hz,2H),0.47(s,2H).HR-MS(ESI):Calculated forC 30 H 35 Br2ClN4O2[M+H] + :677.0888 found 677.0878.

[0321] The synthesis method of compound 60 in Example 8 was adopted to obtain compound 61 by replacing cyclopropyl isocyanate with cyclohexyl isocyanate (cas: 3173-53-3).

[0322]

[0323] 1 H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.3Hz, 1H), 7.57 (s, 1H), 7.50 (d, J = 2.2Hz, 1H), 7.15 (s, 1H), 4.91 (d, J=10.2Hz,1H),4.60(d,J=13.1Hz,1H),4.38(s,1H),3.86(d,J=14.6Hz,3H),3.62(s,2H),3.28(d,J=17 .6Hz,1H),2.99(t,J=15.7Hz,1H),2.81(t,J=13.2Hz,4H),2.38(d,J=12.5Hz,2H),2.22(d,J=5.7Hz,2H ),2.00–1.90(m,3H),1.78–1.68(m,4H),1.44–1.29(m,6H),1.21–1.08(m,6H).HR-MS(ESI):Calculated forC 33 H41 Br2ClN4O2[M+H] + :719.1358 found 719.1355.

[0324] The synthesis method of compound 60 in Example 8 was adopted to obtain compound 62 by replacing cyclopropyl isocyanate with 2-fluorophenyl isocyanate (cas: 16744-98-2).

[0325]

[0326] 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.52 (d, J = 16.7Hz, 2H), 7.27 (s, 1H), 7.25 (s, 1H), 7.14 (s, 1H), 6.99 (t, J = 8.5 Hz,2H),4.95–4.81(m,2H),4.58(d,J=12.8Hz,1H),4.35(d,J=7.3Hz,2H),3.92(d,J=12.2Hz,2H),3.61(t,J=14.4H z,1H),3.27(d,J=17.8Hz,1H),2.90(s,1H),2.82(d,J=14.9Hz,4H),2.43–2.31(m,2H),2.21(d,J=7.3Hz,2H),2.0 0(s,2H),1.74(d,J=10.1Hz,2H),1.53–1.41(m,3H),1.35(s,1H),1.14(d,J=12.3Hz,2H).HR-MS(ESI):Calculated for C 34 H 36 Br2ClFN4O2[M+H] + :745.0950found745.0955.

[0327] The synthesis method of compound 60 in Example 8 was adopted to obtain compound 79 by replacing cyclopropyl isocyanate with isopropyl isocyanate (cas: 1795-48-8).

[0328]

[0329] 1H NMR (300MHz, CDCl3) δ=8.41(s,1H),7.56–7.43(m,2H),7.11(s,1H),4.87(d,J=10.3Hz,1H),4.56(d,J= 13.1Hz,1H),4.26(s,1H),3.82(d,J=17.4Hz,4H),3.58(t,J=13.8Hz,1H),3.23(d,J=17.6Hz,1H),2.95 (t,J=15.7Hz,1H),2.83–2.67(m,4H),2.34(d,J=12.7Hz,2H),2.18(s,2H),1.95(s,1H),1.71(d,J=12. 9Hz,2H),1.31(d,J=10.3Hz,2H),1.23(d,J=9.2Hz,2H),1.11(d,J=6.5Hz,6H).HR-MS(ESI):Calculated forC 30 H 37 Br2ClN4O2[M+H] + :679.1045found679.1040.

[0330] Example 9: Synthesis of (R)-1-(4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidin-1-yl)-3-hydroxypropan-1-one (69)

[0331]

[0332] 3-Hydroxypropionic acid (25.2 mg, 0.14 mmol, cas: 503-66-2), intermediate 48-2 (100 mg, 0.17 mmol) and HATU (80 mg, 0.21 mmol) were dissolved in DMF (5 mL), and DIPEA (71.5 μL) was added after the solution became clear. The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was dropped into ice water, extracted with EA 3 times, washed with water once, washed with brine once, and concentrated under reduced pressure to obtain a yellow oil, which was purified by column chromatography (DCM: MeOH = 40: 1) to obtain 6430 mg of a white solid compound. The yield was 43%. 1H NMR (300MHz, CDCl3) δ=8.41(s,1H),7.56–7.43(m,2H),7.11(s,1H),4.87(d,J=10.3Hz,1H),4.56(d,J= 13.1Hz,1H),4.26(s,1H),3.82(d,J=17.4Hz,4H),3.58(t,J=13.8Hz,1H),3.23(d,J=17.6Hz,1H),2.95 (t,J=15.7Hz,1H),2.83–2.67(m,4H),2.34(d,J=12.7Hz,2H),2.18(s,2H),1.95(s,1H),1.71(d,J=12. 9Hz,2H),1.31(d,J=10.3Hz,2H),1.23(d,J=9.2Hz,2H),1.11(d,J=6.5Hz,6H).HR-MS(ESI):Calculated for C 29 H 34 Br2ClN3O3[M+H] + :666.0728 found 666.0722.

[0333] The synthesis method of compound 69 in Example 9 was adopted to obtain compound 50 by replacing 3-hydroxypropionic acid with glycine (cas: 56-40-6).

[0334]

[0335] 1 H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.3Hz, 1H), 7.60–7.46 (m, 2H), 7.15 (s, 1H), 4.98–4.83 (m, 1H), 4.72– 4.50(m,2H),3.81(s,1H),3.73–3.55(m,2H),3.45(s,2H),3.31–3.23(m,1H),3.02(d,J=13.7Hz,2H),2. 83(d,J=14.2Hz,2H),2.63(t,J=13.3Hz,1H),2.41(t,J=12.6Hz,2H),2.29–2.16(m,2H),2.16–2.04(m, 1H),1.61–1.45(m,2H),1.44–1.32(m,2H),1.27(d,J=9.4Hz,2H),1.13(s,2H).HR-MS(ESI):Calculated forC 28 H 33 Br2ClN4O2[M+H] +:651.0732found651.0730.

[0336] The synthesis method of compound 69 in Example 9 was adopted to obtain compound 85 by replacing 3-hydroxypropionic acid with 3-(methylsulfonyl)propionic acid (cas: 645-83-0).

[0337]

[0338] 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.55 (s, 1H), 7.50 (s, 1H), 7.15 (s, 1H), 4.90 (d, J = 10.2Hz, 1H), 4.58 (s, 2H), 3 .81(s,2H),3.62(t,J=14.0Hz,1H),3.42(d,J=7.5Hz,2H),3.27(d,J=17.3Hz,1H),3.15–3.00(m,2H),2.98(s,3H), 2.92–2.84(m,3H),2.81(d,J=5.1Hz,1H),2.63(t,J=13.1Hz,1H),2.41(t,J=12.9Hz,2H),2.22(t,J=5.8Hz,2H),2 .10(d,J=15.3Hz,1H),1.90–1.72(m,4H),1.52(d,J=13.6Hz,2H),1.39(d,J=16.3Hz,2H).HR-MS(ESI):Calculated for C 30 H 36 Br2ClN3O4S[M+H] + :728.0555found728.0550.

[0339] The synthesis method of compound 69 in Example 9 was adopted to obtain compound 86 by replacing 3-hydroxypropionic acid with 4,4,4-trifluorobutyric acid (cas: 406-93-9).

[0340]

[0341] 1H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.55 (s, 1H), 7.52–7.47 (m, 1H), 7.15 (s, 1H), 4.90 (d, J = 10.3Hz, 1H), 4.60 (d,J=12.9Hz,2H),3.83(d,J=12.3Hz,2H),3.62(t,J=13.9Hz,1H),3.27(d,J=18.2Hz,1H),3.05(q,J=12.8Hz, 2H),2.91–2.80(m,2H),2.56(t,J=10.7Hz,6H),2.47–2.34(m,2H),2.29–2.17(m,2H),2.12(s,1H),1.88(d,J= 13.0Hz,1H),1.78(d,J=13.1Hz,2H),1.52(d,J=13.7Hz,2H),1.40(d,J=15.3Hz,2H).HR-MS(ESI):Calculated for C 30 H 33 Br2ClF3N3O2[M+H] + :718.0653 found 718.0650.

[0342] The synthesis method of compound 69 in Example 9 was adopted to obtain compound 87 by replacing 3-hydroxypropionic acid with 3-cyanopropionic acid (cas: 16051-87-9).

[0343]

[0344] 1 H NMR (300MHz, CDCl3) δ=δ=8.44(s,1H),7.55(s,1H),7.52–7.47(m,1H),7.15(s,1H),4.90(d,J=10.3Hz,1H),4. 60(d,J=12.9Hz,2H),3.83(d,J=12.3Hz,2H),3.62(t,J=13.9Hz,1H),3.27(d,J=18.2Hz,1H),3.05(q,J=12.8Hz ,2H),2.91–2.80(m,2H),2.96(t,J=10.7Hz,6H),2.47–2.34(m,2H),2.29–2.17(m,2H),2.12(s,1H),1.88(d,J =13.0Hz,1H),1.78(d,J=13.1Hz,2H),1.52(d,J=13.7Hz,2H),1.40(d,J=15.3Hz,2H).HR-MS(ESI):Calculated for C30 H 33 Br2ClN4O2[M+H] + :675.0732 found 675.0730.

[0345] Example 10: Synthesis of ((R)-4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptane[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)-N-(2-hydroxyethyl)piperidine-1-carboxamide (72)

[0346]

[0347] The synthesis of intermediate 48-2 is the same as that in Example 5.

[0348] Synthesis of intermediate 72-1:

[0349] 2-(tert-Butyldimethylsilyl)oxy)ethane-1-amine (35.3 mg, 0.20 mmol, cas: 17348-66-2) was dissolved in anhydrous DCM (5 mL), triphosgene (19.9 mg, 0.06 mmol, cas: 32315-10-9) and TEA (71 μL) were added under ice bath, and the mixture was reacted at room temperature for 3 h. Subsequently, intermediate 48-2 (100 mg, 0.17 mmol) and TEA (47 μL) were added, and the mixture was reacted overnight. After the reaction was completed as monitored by TLC, saturated NH4Cl aqueous solution was added to quench the mixture, and the mixture was extracted with DCM three times, washed with water three times, washed with brine once, and concentrated under reduced pressure to obtain light yellow foamy solid intermediate 76-1100 mg, which was directly reacted in the next step without purification.

[0350] Synthesis of compound 72:

[0351] 76-1 (100 mg, 0.20 mmol) was dissolved in anhydrous THF (5 mL), TBAF (85 mg, 0.26 mmol) was added dropwise in an ice bath, and then reacted at room temperature for 2 h. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure to remove THF, and water (10 mL) was added to the residue, extracted three times with EA, washed three times with water, washed once with brine, concentrated under reduced pressure, and purified by column chromatography (DCM: MeOH = 20: 1) to obtain 7680 mg of a white solid compound. The yield was 83%. 1H NMR (300MHz, CDCl3) δ=8.47(d,J=8.1Hz,1H),7.61(d,J=18.6Hz,1H),7.51(d,J=2.2Hz,1H),7.16(s,1H),4.95(dd,J=10.2,7.1Hz ,1H),4.61(d,J=13.3Hz,1H),3.99(d,J=15.1Hz,1H),3.91(s,1H),3.87–3.67(m,3H),3.62(s,1H),3.41(d,J=5.4Hz,2H),3.35–3 .22(m,1H),3.03(d,J=15.4Hz,1H),2.92–2.84(m,2H),2.84–2.75(m,2H),2.61(s,1H),2.38(d,J=11.8Hz,2H),2.28–2.14(m,2H) ,2.03(s,1H),1.77(d,J=11.4Hz,2H),1.51(d,J=13.0Hz,2H),1.45–1.33(m,2H),1.17(d,J=11.6Hz,2H).HR-MS(ESI):Calculated for C 29 H 35 Br2ClN4O3[M+H] + :681.0837 found 681.0830.

[0352] The synthesis method of compound 72 in Example 10 was used to replace 2-(tert-butyldimethylsilyl)oxy)ethane-1-amine with deuterated methylamine hydrochloride (cas: 7436-22-8) to obtain compound 59.

[0353]

[0354] 1H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H) ,7.30(s,1H),7.00–7.95(m,1H),3.92(d,J=9.5Hz,1H),3.72–3.66(m,2H),3.59–3.54(m,4 H),3.31–3.25(m,2H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.32–2.26(m,1H),2.28–2.16 (m,2H),2.07–1.97(m,1H),1.92–1.78(m,4H),1.82–1.65(m,4H).HR-MS(ESI):Calculated for C 28 H 30 D3Br2ClN4O2[M+H] + :654.0920 found 654.0915.

[0355] The synthesis method of compound 72 in Example 10 was adopted to replace 2-(tert-butyldimethylsilyl)oxy)ethane-1-amine with O-(tetrahydro-2H-pyran)-2-hydroxylamine (cas: 6723-30-4) to obtain compound 70.

[0356]

[0357] 1 H NMR (300MHz, CDCl3) δ = 8.45 (s, 1H), 7.53 (dd, J = 19.9, 4.3Hz, 2H), 7.15 (s, 1H), 4.91 (d, J = 10 .0Hz,1H),4.60(s,1H),3.89(d,J=41.1Hz,3H),3.60(d,J=14.4Hz,1H),3.28(d,J=17.5Hz,1 H),3.01(d,J=14.3Hz,1H),2.83(d,J=14.2Hz,4H),2.39(s,2H),2.22(s,2H),2.05(s,1H),1 .90–1.69(m,2H),1.52(s,2H),1.40(d,J=16.4Hz,3H),1.18(s,2H).HR-MS(ESI):Calculated for C 27 H 31 Br2ClN4O3[M+H] + :653.0524found653.0520.

[0358] The synthesis method of compound 72 in Example 10 was used to replace 2-(tert-butyldimethylsilyl)oxy)ethyl-1-amine with (S)-2-((tert-butyldimethylsilyl)oxy)butane-1-amine to obtain compound 73

[0359]

[0360] 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(dt,J=1.9,0.9Hz ,1H),5.81(t,J=7.0Hz,1H),3.92(d,J=9.5Hz,1H),3.74–3.50(m,7H),3.49(d,J=5.7Hz,1H),3.35–3.29(m,1H),3. 32–3.22(m,2H),3.21–3.15(m,1H),3.07–2.98(m,1H),3.00–2.90(m,3H),2.32(dd,J=15.1,6.9Hz,1H),2.28–2.1 6(m,2H),2.07–1.97(m,1H),1.92–1.65(m,8H),1.47–1.35(m,2H),0.94(t,J=7.2Hz,3H).HR-MS(ESI):Calculated for C 31 H 39 Br2ClN4O3[M+H] + :709.1150 found 709.1140.

[0361] The synthesis method of compound 72 in Example 10 was adopted to replace 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid to obtain compound 74.

[0362]

[0363] 1H NMR (300MHz, CDCl3) δ = 8.45 (s, 1H), 7.53 (d, J = 16.0Hz, 2H), 7.16 (s, 1H), 5.09 (t, J = 5.6Hz, 1H), 4.93 (s,1H),4.89(s,1H),4.53(d,J=11.8Hz,1H),4.02(d,J=12.8Hz,3H),3.71(s,2H),3.59(d,J=12.3Hz ,2H),3.46–3.35(m,2H),3.34–3.22(m,1H),3.07–2.88(m,2H),2.81(s,3H),2.56–2.33(m,2H),2.11 (s,1H),1.84(d,J=15.7Hz,4H),1.59(d,J=17.4Hz,2H),1.45–1.35(m,4H).HR-MS(ESI):Calculated for C 29 H 34 Br2ClFN4O3[M+H] + :699.0743 found 699.0740.

[0364] The synthesis method of compound 72 in Example 10 was used, 1-tert-butoxycarbonyl-4-piperidineacetic acid was replaced by 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid, and 2-(tert-butyldimethylsilyl)oxy)ethane-1-amine was replaced by 1-aminomethyl-1-cyclopropanol (cas: 74592-33-9) to obtain compound 75.

[0365]

[0366] 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.59–7.47 (m, 2H), 7.16 (s, 1H), 4.91 (d, J = 10.4Hz, 1H), 4.80–4 .72(m,2H),4.55(s,1H),4.03(d,J=13.4Hz,3H),3.62(t,J=13.8Hz,1H),3.34(s,2H),3.27(d,J=16 .9Hz,1H),3.06–2.89(m,2H),2.89–2.76(m,3H),2.47(d,J=23.0Hz,2H),2.12(s,1H),1.91–1.80(m ,1H),1.69–1.51(m,3H),1.40(d,J=12.7Hz,4H),0.80(s,2H),0.54(s,2H).HR-MS(ESI):Calculated for C 31H 36 Br2ClFN4O3[M+H] + :725.0899 found 725.0889.

[0367] The synthesis method of compound 72 in Example 10 was adopted to replace 2-(tert-butyldimethylsilyl)oxy)ethane-1-amine with 1-aminocyclobutanemethanol hydrochloride (cas: 1392213-15-8) to obtain compound 76.

[0368]

[0369] 1 H NMR (300MHz, CDCl3) δ = 8.46 (s, 1H), 7.58 (d, J = 9.3Hz, 1H), 7.51 (t, J = 2.8Hz, 1H), 7.16 (s, 1H), 4.94 (t, J = 5.1Hz, 1H), 4.87 (d,J=10.7Hz,1H),4.61(d,J=13.2Hz,1H),3.88(d,J=15.0Hz,3H),3.77(d,J=3.7Hz,2H),3.63(t,J=15.4Hz,1H),3.29(d, J=17.5Hz,1H),3.02(d,J=14.5Hz,1H),2.92–2.74(m,4H),2.39(d,J=12.2Hz,2H),2.30–2.18(m,4H),2.11–1.95(m,4H),1 .91(s,1H),1.82–1.70(m,2H),1.51(d,J=12.7Hz,2H),1.40(d,J=16.4Hz,2H),1.25–1.06(m,2H).HR-MS(ESI):Calculated for C 32 H 39 Br2ClN4O3[M+H] + :721.1150 found721.1140.

[0370] The synthesis method of compound 72 in Example 10 was adopted to replace 2-(tert-butyldimethylsilyl)oxy)ethane-1-amine with -(aminomethyl)cyclobutanol (cas: 180205-28-1) to obtain compound 77.

[0371]

[0372] 1H NMR (300MHz, CDCl3) δ==8.44(d,J=2.6Hz,1H),7.60–7.47(m,2H),7.28(s,1H),7.15(s,1H),5.09(s,1H),4.89(dd,J=10.2,3.1Hz, 1H),4.60(d,J=13.5Hz,1H),3.92(d,J=11.8Hz,2H),3.83(s,1H),3.70–3.55(m,1H),3.39(d,J=5.3Hz,2H),3.32–3.21(m,1H),3.0 1(d,J=15.2Hz,1H),2.90–2.75(m,4H),2.39(d,J=12.9Hz,2H),2.28–2.19(m,2H),2.06(d,J=9.0Hz,3H),2.01(s,2H),1.82(d,J=1 6.0Hz,2H),1.71(d,J=20.6Hz,2H),1.54(s,1H),1.51–1.38(m,3H),1.38–1.31(m,1H),1.23–1.08(m,2H).HR-MS(ESI):Calculated for C 32 H 39 Br2ClN4O3[M+H] + :721.1150 found 721.1140.

[0373] The synthesis method of compound 72 in Example 10 was used to replace 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid and 2-(tert-butyldimethylsilyl)oxy)ethyl-1-amine with 2-methoxyethylamine (cas: 109-85-3) to obtain compound 78.

[0374]

[0375] 1H NMR (300MHz, CDCl3) δ = 8.45 (s, 1H), 7.54 (d, J = 19.3Hz, 2H), 7.15 (s, 1H), 4.93 (d, J = 10.1Hz, 2H), 4.87 (s, 1H), 4.6 2–4.50(m,1H),4.01(d,J=8.3Hz,2H),3.98(s,1H),3.62(t,J=14.1Hz,1H),3.47(d,J=4.8Hz,2H),3.42(d,J=5.8H z,2H),3.36(s,3H),3.27(d,J=17.9Hz,1H),3.08–2.89(m,2H),2.88–2.83(m,1H),2.83–2.72(m,2H),2.46(d,J=1 9.8Hz,2H),2.11(s,1H),1.88(s,2H),1.65(s,1H),1.58(d,J=14.2Hz,2H),1.41(s,4H).HR-MS(ESI):Calculated for C 30 H 36 Br2ClFN4O3[M+H] + :713.0899 found 713.0890.

[0376] Example 11: Synthesis of (R)-N-(2-aminoethyl)-4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidine-1-carboxamide (80)

[0377]

[0378] The synthesis of intermediate 48-1 was the same as in Example 5.

[0379] Synthesis of intermediate 80-1:

[0380] N-tert-Butyloxycarbonyl-1,2-ethylenediamine (30.3 mg, 0.20 mmol, cas: 57260-73-8) was dissolved in anhydrous DCM (5 mL), triphosgene (19.9 mg, 0.06 mmol, cas: 32315-10-9) and TEA (65 μL) were added under ice bath, and the mixture was reacted at room temperature for 3 h. Then, intermediate 48-2 (100 mg, 0.17 mmol) and TEA (47 μL) were added, and the mixture was reacted overnight at room temperature. After the reaction was completed as monitored by TLC, saturated NH4Cl aqueous solution was added to quench the reaction, and the mixture was extracted with DCM three times, washed with water three times, washed with brine once, and concentrated under reduced pressure to obtain light yellow foamy solid intermediate 84-1100 mg, which was directly reacted in the next step without purification.

[0381] Synthesis of compound 80:

[0382] 84-1 (100 mg, 0.20 mmol) was dissolved in EA (5 mL), and HCl-EA (0.5 mL) was added dropwise under ice bath, followed by reaction at room temperature for 2 h. After the reaction was completed as monitored by TLC, the mixture was concentrated under reduced pressure to remove EA, and water (10 mL) and 25% ammonia water (0.5 mL) were added to the residue, washed with water three times and brine once, concentrated under reduced pressure, and purified by column chromatography (DCM:MeOH=20:1) to obtain 8080 mg of a white solid compound. The yield was 83%. 1 H NMR (300MHz, CDCl3) δ = 8.44 (s, 1H), 7.53 (d, J = 15.6Hz, 2H), 7.15 (s, 1H), 5.25 (s, 1H), 4.90–4.8 5(m,1H),4.60–4.55(m,1H),3.92(d,J=9.5Hz,1H),3.69–3.64(m,2H),3.58–3.43(m,4H),3.35– 3.30(m,2H),3.27–3.24(m,2H),3.10–3.03(m,2H),3.06–2.98(m,1H),3.00–2.90(m,3H),2.37– 2.33(m,1H),2.28–2.16(m,2H),2.07–1.97(m,1H),1.92–1.65(m,8H).HR-MS(ESI):Calculated for C 29 H 36 Br2ClN5O2[M+H] + :680.0997 found 680.0990.

[0383] The synthesis method of compound 80 in Example 11 was used to replace N-tert-butyloxycarbonyl-1,2-ethylenediamine with tert-butyl n-[1-(aminomethyl)cyclobutyl]carbamate (cas: 1286330-22-0) to obtain compound 81.

[0384]

[0385] 1H NMR (300MHz, CDCl3) δ = 8.45 (s, 1H), 7.61–7.48 (m, 2H), 7.15 (s, 1H), 5.18 (t, J = 5.0Hz, 1H), 4.90 (dd, J = 10.4, 2.6Hz, 1H), 4 .59(d,J=13.0Hz,1H),3.96(d,J=12.8Hz,2H),3.83(t,J=11.9Hz,1H),3.62(t,J=14.2Hz,1H),3.34(d,J=5.0Hz,2H),3.31 –3.21(m,1H),2.98(t,J=19.4Hz,1H),2.91–2.74(m,4H),2.45–2.31(m,2H),2.25–2.20(m,2H),2.02–1.29(d,J=9.4Hz,2H ),1.89–1.79(m,2H),1.79–1.70(m,4H),1.59–1.40(m,3H),1.39–1.29(m,2H),1.21–1.10(m,2H).HR-MS(ESI):Calculated for C 32 H 40 Br2ClN5O2[M+H] + :720.1310 found 720.1303.

[0386] The synthesis method of compound 80 in Example 11 was used to replace N-tert-butyloxycarbonyl-1,2-ethylenediamine with (1-amino-2-methylpropan-2-yl)carbamic acid tert-butyl ester (cas: 320581-09-7) to obtain compound 82.

[0387]

[0388] 1H NMR (300MHz, CDCl3) δ = 8.45 (s, 1H), 7.53 (d, J = 15.4Hz, 2H), 7.15 (s, 1H), 5.15 (s, 1H), 4.90 (d, J = 10.1Hz, 1H), 4. 60(d,J=10.9Hz,1H),3.96(d,J=12.6Hz,2H),3.83(s,1H),3.62(t,J=14.3Hz,1H),3.27(d,J=17.7Hz,1H),3.14( d,J=5.4Hz,2H),3.01(d,J=15.2Hz,1H),2.83(d,J=14.7Hz,4H),2.38(d,J=12.8Hz,2H),2.27–2.17(m,2H),2.01 (s,1H),1.73(s,2H),1.47(dd,J=24.8,12.0Hz,3H),1.35(d,J=8.8Hz,1H),1.13(s,8H).HR-MS(ESI):Calculated for C 31 H 40 Br2ClN5O2[M+H] + :708.1310 found 708.1304.

[0389] The synthesis method of compound 80 in Example 11 was used to replace 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid and N-tert-butoxycarbonyl-1,2-ethylenediamine with N1,N1-diethylbutane-1,4-diamine (cas: 27431-62-5) to obtain compound 83.

[0390]

[0391] 1H NMR (300MHz, CDCl3) δ = 8.45 (d, J = 2.3Hz, 1H), 7.60–7.47 (m, 2H), 7.15 (s, 1H), 5.01 (t, J = 5.2Hz, 1H), 4.91 (d,J=10.2Hz,1H),4.55(s,1H),3.98(d,J=11.4Hz,3H),3.62(t,J=13.8Hz,1H),3.33–3.17(m,3H),3.06– 2.68(m,5H),2.58–2.49(m,4H),2.42(d,J=6.4Hz,3H),2.09(s,1H),1.94(s,3H),1.87(s,1H),1.67–1.54 (m,3H),1.38(q,J=9.4,8.7Hz,4H),1.27(d,J=8.9Hz,2H),1.02(t,J=7.2Hz,6H).HR-MS(ESI):Calculated forC 35 H 47 Br2ClFN5O2[M+H] + :782.1842 found 782.1822.

[0392] The synthesis method of compound 80 in Example 11 was used to replace 1-tert-butoxycarbonyl-4-piperidineacetic acid with 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-fluoroacetic acid and N-tert-butoxycarbonyl-1,2-ethylenediamine with tert-butyl (2-(2-aminoethoxy)ethyl)carbamate (cas: 127828-22-2) to obtain compound 84.

[0393]

[0394] 1H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s ,1H),5.75(t,J=5.6Hz,1H),5.11(d,J=6.4Hz,0H),3.92(d,J=9.5Hz,1H),3.74–3.49(m,9H),3.41 –3.35(m,2H),3.33–3.27(m,2H),3.07–2.96(m,1H),2.99–2.90(m,3H),2.85–2.75(m,2H),2.43–2 .37(m,1H),2.30(d,J=14.1Hz,1H),2.21–2.16(m,1H),1.85–1.67(m,8H).HR-MS(ESI):Calculated for C 31 H 39 Br2ClFN5O3[M+H] + :742.1165 found 742.1145.

[0395] Example 12: Synthesis of (R)-4-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)ethyl)piperidine-1-carboxamide (39)

[0396]

[0397] Synthesis of 4-(2-bromoethyl)piperidine-1-carboxamide:

[0398]

[0399] 4-(2-Bromoethyl)piperidine hydrobromide (0.5 g, cas: 69712-10-3) was added to a single-mouth bottle, and a mixed solvent of N-methyl-2-pyrrolidone NMP:H2O (4:1, 5 mL) was added, followed by urea (2.5 g). The mixture was refluxed at 130°C for 9 h. After the TLC reaction was complete, the temperature was lowered to 25°C, and the reaction solution was poured into water, extracted with DCM three times, washed with water three times, washed with brine once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a colorless oil, which was directly used for the next step without purification.

[0400] Synthesis of compound 39:

[0401] The intermediate IX (0.5 g, 1.06 mmol) was dissolved in 5 mL of acetonitrile, and 4-(2-bromoethyl)piperidine-1-carboxamide (0.27 g, 1.17 mmol) and potassium carbonate (0.29 g, 2.12 mmol) were added, followed by reaction at 75°C for 2 h. After the TLC reaction was complete, water was added, and DCM was extracted three times. The organic layers were combined, washed with water and brine once, concentrated, and purified by column chromatography (DCM:MeOH=50:1) to obtain 390.51 g of a white solid compound. 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6Hz, 1H), 7.78 (d, J = 1.6Hz, 1H), 7.47 (d, J = 2.2Hz, 1H), 7. 00–76.95(m,1H),5.29(s,2H),3.92(d,J=9.5Hz,1H),3.67–3.59(m,2H),3.57–3.50(m,2H),3. 07–2.94(m,2H),2.98–2.90(m,2H),2.71–2.65(m,2H),2.46–2.40(m,1H),2.40–2.34(m,1H),2 .37–2.26(m,2H),2.18(m,1H),1.77–1.57(m,9H),1.47–1.41(m,2H).HR-MS(ESI):Calculated for C 27 H 33 Br2ClN4O[M+H] + :623.0782 found 623.0775.

[0402] The synthesis method of compound 39 in Example 12 was adopted to obtain compound 38 by replacing 4-(2-bromoethyl)piperidine-1-carboxamide with 4-(2-chloroacetyl)piperazine-1-carboxamide.

[0403]

[0404] Synthesis of 4-(2-chloroacetyl)piperazine-1-carboxamide:

[0405]

[0406] Piperazinecarboxamide hydrochloride (250 mmol) was dispersed in DCM (5 mL), and then chloroacetyl chloride (300 mmol) and triethylamine (600 mmol) were added dropwise. The mixture was reacted at room temperature for 2 h. A saturated aqueous sodium bicarbonate solution (10 mL) was added to quench the reaction. The mixture was extracted with DCM three times, washed once with water and once with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a colorless oil, which was directly used for the next step without purification.

[0407] Synthesis of compound 38:

[0408] 1 H NMR (300MHz, CDCl3) δ = 8.44 (d, J = 1.6Hz, 1H), 7.78 (d, J = 1.6Hz, 1H), 7.47 (d, J = 2.2Hz, 1H), 7.00–6. 97(m,1H),5.30(d,J=7.0Hz,1H),5.24(d,J=6.9Hz,1H),3.92(d,J=9.5Hz,1H),3.69–3.61(m,2H),3. 61–3.55(m,4H),3.55–3.47(m,2H),3.15(s,2H),3.07–2.95(m,2H),2.98–2.92(m,2H),2.95–2.90(m ,1H),2.90–2.85(m,1H),2.43(m,2H),2.18–2.14(m,1H),1.73–1.58(m,4H).HR-MS(ESI):Calculate d for C 26 H 30 Br2ClN5O2[M+H] + :638.0528 found 638.0525.

[0409] The synthesis method of compound 39 in Example 12 was adopted to obtain compound 40 by replacing 4-(2-bromoethyl)piperidine-1-carboxamide with 4-(bromomethyl)piperidine-1-carboxamide.

[0410]

[0411] Synthesis of 4-(2-bromomethyl)piperidine-1-carboxamide:

[0412]

[0413] 4-(2-Bromomethyl)piperidinium hydrogen hydrochloride (0.5 g, cas: 1159825-22-5) was added to a single-mouth bottle, and a mixed solvent of N-methyl-2-pyrrolidone NMP:H2O (4:1, 5 mL) was added, followed by urea (2.5 g). The mixture was refluxed at 130°C for 9 h. After the TLC reaction was complete, the temperature was lowered to 25°C, and the reaction solution was poured into water, extracted with DCM three times, washed with water three times, washed with brine once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a colorless oil, which was directly used for the next step without purification.

[0414] Synthesis of compound 40:

[0415] 1H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H ),7.00–7.95(m,1H),5.29(s,2H),3.92(d,J=9.5Hz,1H),3.67–3.62(m,2H),3.58–3.53(m ,2H),3.07–2.94(m,2H),2.98–2.90(m,2H),2.74–2.65(m,2H),2.61–2.56(m,1H),2.54– 2.50(m,1H),2.40–2.31(m,2H),2.18(m,1H),1.81–1.61(m,9H).HR-MS(ESI):Calculated for C 26 H 31 Br2ClN4O[M+H] + :609.0620 found 609.0610.

[0416] Example 13: Synthesis of 4-(2-(4-(8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidine-1-carboxamide (11)

[0417]

[0418] Synthesis of intermediate 11-2:

[0419] The raw material 11-18-chloro-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-one (5 g, 20.5 mmol, cas: 31251-41-9) was dissolved in 60 mL of acetic acid-trifluoroacetic acid mixed solvent (volume ratio 5:1), and then zinc powder (3.99 g, 61.5 mmol) was added. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by TLC, the reaction solution was poured into a saturated aqueous sodium carbonate solution, and the solid was precipitated. The solid was filtered and dried to obtain 4.3 g of a yellow solid, which was directly used for the next step without purification.

[0420] Synthesis of intermediate 11-3:

[0421] 1-Boc-4-methanesulfonyloxypiperidine (5.48 g, 20.9 mmol) and intermediate 11-2 (4.0 g, 17.41 mmol) were dissolved in 15 mL of toluene, cooled to 25°C, and 15 mL of lithium diisopropylamide LDA was slowly added. After 4 hours, the reaction was complete as monitored by TLC. Water was added to quench the reaction, and the organic layer was separated. The aqueous layer was extracted with ethyl acetate, washed with water and saturated brine, dried, concentrated, and then column chromatography (PE:EA=10:1) was performed to obtain a white solid intermediate 11-33 g.

[0422] Synthesis of intermediate 11-4:

[0423] 2 g of intermediate 11-3 was dissolved in 5 mL of ethyl acetate, and 10 mL of hydrogen chloride / ethyl acetate solution was added. The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, it was filtered and dried to obtain a white solid intermediate 11-4.

[0424] Synthesis of compound 11:

[0425] The intermediate 11-4 (1 g, 3.20 mmol), 2-(1-carbamoylpiperidin-4-yl)acetic acid (0.63 g, 3.36 mmol, cas: 279236-52-1), EDCI (0.92 g, 4.80 mmol) and HOBT (0.65 g, 4.80 mmol) were dissolved in DMF (20 mL), stirred at room temperature for 30 min, and TEA (0.90 g, 9.60 mmol) was added dropwise. The mixture was reacted at room temperature overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water. Solids precipitated and then filtered. After the filter cake was dried, it was purified by silica gel column chromatography (DCM: MeOH = 50: 1) to obtain the intermediate 23-31.3 g with a yield of 84.3%. 1H NMR (300MHz, CDCl3) δ = 8.39 (dd, J = 4.7, 1.9 Hz, 1H), 7.43 ( dd, J = 8.1, 1.8 Hz, 1H), 7.27 ( dd, J = 8.4, 2.2 Hz, 1H), 7.23 –7.16(m,2H),7.06–7.01(m,1H),5.29(s,2H),4.24–4.18(m,1H),3.76–3.66(m,4H),3.66–3.61(m,1H),3.61(dd, J=8.4,6.6Hz,1H),3.32–3.28(m,2H),3.05–2.95(m,1H),2.88–2.85(m,1H),2.84–2.77(m,2H),2.36–2.29(m,1H) ,2.32–2.26(m,1H),2.25(dd,J=15.2,7.0Hz,1H),2.01–1.97(m,1H),1.88–1.71(m,9H).HR-MS(ESI):Calculated for C 27 H 33 ClN4O2[M+H] + :481.2365 found 481.2360.

[0426] The synthesis method of compound 11 in Example 13 was adopted to obtain compound 12 by replacing 11-1 with 3-bromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridine.

[0427]

[0428] 1H NMR (300MHz, CDCl3) δ=8.39(dd,J=4.7,1.9Hz,1H),,7.27(dd,J=8.4,2.2Hz,1H),7.23–7.16(m,2H),7.06– 7.01(m,1H),5.29(s,2H),4.24–4.18(m,1H),3.76–3.66(m,4H),3.66–3.61(m,1H),3.61(dd,J=8.4,6.6Hz ,1H),3.32–3.28(m,2H),3.05–2.95(m,1H),2.88–2.85(m,1H),2.84–2.77(m,2H),2.36–2.29(m,1H),2.32 –2.26(m,1H),2.25(dd,J=15.2,7.0Hz,1H),2.01–1.97(m,1H),1.88–1.71(m,9H).HR-MS(ESI):Calculated forC 27 H 32 BrClN4O2[M+H] + :559.1470 found 559.1465.

[0429] The synthesis method of compound 11 in Example 13 was adopted to obtain compound 13 by replacing 11-1 with 10-bromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridine.

[0430]

[0431] 1H NMR (300MHz, CDCl3) δ = 8.39 (dd, J = 4.7, 1.9 Hz, 1H), 7.43 ( dd, J = 8.1, 1.8 Hz, 1H), 7.27 ( dd, J = 8.4, 2.2 Hz, 1H), 7.23 –7.16(m,1H),7.06–7.01(m,1H),5.29(s,2H),4.24–4.18(m,1H),3.76–3.66(m,4H),3.66–3.61(m,1H),3.61(dd, J=8.4,6.6Hz,1H),3.32–3.28(m,2H),3.05–2.95(m,1H),2.88–2.85(m,1H),2.84–2.77(m,2H),2.36–2.29(m,1H) ,2.32–2.26(m,1H),2.25(dd,J=15.2,7.0Hz,1H),2.01–1.97(m,1H),1.88–1.71(m,9H).HR-MS(ESI):Calculated for C 27 H 32 BrClN4O2[M+H] + :559.1470 found 559.1463.

[0432] Example 14: Synthesis of 4-(2-(6-(8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)-2-aza[3.3]heptane-2-yl)-2-oxoethyl)piperidine-1-carboxamide (7)

[0433]

[0434] Compound 5 (1 g, 2.04 mmol) was dissolved in 10 mL of toluene, and the temperature was raised to 120°C after nitrogen replacement, and 4.6 mL of DIBAL was added dropwise, followed by reaction for 2 h. After the reaction was complete as monitored by HPLC, the temperature was lowered to room temperature, 5 mL of water was added dropwise to quench the reaction, and then EA was extracted 3 times. The organic layers were combined, washed once with water, once with brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The compound 70.5 g was obtained by purification by silica gel column chromatography (DCM: MeOH = 50: 1). 1H NMR (300MHz, CDCl3) δ=8.39 (dd, J=4.7, 1.9Hz, 1H), 7.43 (dd, J=8.1, 1.8Hz, 1H), 7.27 (dd, J=8.4 ,2.2Hz,1H),7.23–7.16(m,2H),7.06(dt,J=2.2,1.0Hz,1H),5.29(s,2H),4.27(d,J=7.9Hz,1H) ,3.74–3.57(m,8H),3.05–2.95(m,1H),2.88–2.83(m,1H),2.84–2.77(m,2H),2.40–2.34(m,1H) ,2.28(d,J=7.0Hz,2H),2.06–1.97(m,1H),1.90–1.74(m,6H),1.69–1.60(m,2H).HR-MS(ESI):Ca lculatedfor C 28 H 33 ClN4O2[M+H] + :493.2365 found 493.2361.

[0435] Example 15: Synthesis of (R)-N-(2-aminoethyl)-1-(2-(4-(3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-yl)piperidin-1-yl)-2-oxoethyl)piperidine-4-carboxamide (51)

[0436]

[0437] Synthesis of intermediate 51-1:

[0438] 14-1 (50 mmol) was dissolved in acetonitrile (5 mL), followed by the addition of methyl piperidine-4-carboxylate (60 mmol) and potassium carbonate (100 mmol), and the mixture was reacted at 70°C for 2 h, and 5 mL of water was added to quench the reaction, followed by extraction with EA for 3 times. The organic layers were combined, washed once with water and once with brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure, and the mixture was directly used for the next step.

[0439] Synthesis of intermediate 51-2:

[0440] 51-1 (0.2 g) was dissolved in ethanol (5 mL), and then 1.5 N NaOH aqueous solution (2 mL) was added dropwise. After reacting at room temperature for 3 h, the solvent was removed under reduced pressure, 1 N HCl (3 mL) was added dropwise to adjust the pH to 4, and extracted with DCM three times. The organic layers were combined, washed once with brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure, and the mixture was directly used for the next step.

[0441] Synthesis of intermediate 51-3:

[0442] Intermediate 51-2 (0.2 g), N-tert-butyloxycarbonyl-1,2-ethylenediamine (0.08 g, cas: 279236-52-1), EDCI (0.44 g) and HOBT (0.31 g) were dissolved in DMF (5 mL), stirred at room temperature for 30 min, and TEA (0.27 g) was added dropwise. The mixture was allowed to react overnight at room temperature. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water, and solid precipitated. The solid was then filtered off. After the filter cake was dried, it was purified by silica gel column chromatography (DCM: MeOH = 50: 1) to obtain intermediate 23-30.15 g.

[0443] Synthesis of intermediate 51:

[0444] 51-3 (150 mg, 0.20 mmol) was dissolved in EA (5 mL), and HCl-EA (0.7 mL) was added dropwise under an ice bath. The mixture was then reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the mixture was concentrated under reduced pressure to remove EA. Water (5 mL) and 25% aqueous ammonia (0.5 mL) were added to the residue. The mixture was washed three times with water and once with brine. After concentration under reduced pressure, the mixture was purified by column chromatography (DCM:MeOH=20:1) to obtain 5180 mg of a white solid compound. 1 H NMR (300MHz, CDCl3) δ=8.44(d,J=1.6Hz,1H),7.78(d,J=1.6Hz,1H),7.47(d,J=2.2Hz,1H),7.00(s,1H),6.8 9(t,J=5.1Hz,1H),4.08(t,J=6.3Hz,2H),3.95–3.84(m,3H),3.35–3.31(m,1H),3.34–3.29(m,1H),3.31–3.2 6(m,1H),3.29–3.23(m,1H),3.17(d,J=13.2Hz,1H),3.11(d,J=13.0Hz,1H),3.07–2.90(m,6H),2.88–2.80( m,2H),2.51–2.46(m,2H),2.37–2.32(m,1H),2.21–2.17(m,1H),1.89–1.64(m,8H).HR-MS(ESI):Calculated for C 29 H 36 Br2ClN5O2[M+H] + :680.0997 found 680.0992.

[0445] Example 16: Inhibitory activity of the compounds of the present invention against RSV

[0446] 1. Experimental Methods

[0447] Virus cytopathic effect (CPE) assay: 96-well plates were plated with 1.3 × 10 4 Hep-2 cells were seeded at a density of , and the cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS). The next day, the cells were infected with RSV at an MOI of 0.2, which produced a cytopathic effect of about 90% after 5 days, with a total volume of 200 μL per well. During this period, the cells were incubated in the presence or absence of serial dilutions of the compound. The viability of the cells was assessed by CCK-8 (Dojindo Molecular Technologies, Inc.) after 5 days. The absorbance at 450 nm and the reference value at 630 nm were measured to determine the 50% effective concentration (EC 50 ).

[0448] 2. Experimental Results

[0449] Table 1. Cellular activities of the compounds of the present invention

[0450]

[0451]

[0452]

[0453] As can be seen from Table 1, the compounds of the present invention have inhibitory activity against RSV, which is significantly improved compared with the positive control. Therefore, the compounds of the present invention can be used as small molecule RSV inhibitors.

[0454] Example 17: PK study of the compounds of the present invention in mice

[0455] 1. Experimental Methods

[0456] The compounds used in this application were prepared using a DMSO: PEG-400: hydroxypropyl-β-cyclodextrin (20:40:40, v / v / v) system and administered orally (PO) to fasting CD-1 mice (male, 10–12 g, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.) at a dose of 25 mg of the compound per kg body weight. Plasma and lung samples were collected 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, and 24 h after administration. The concentration of the compound was determined by liquid chromatography-mass spectrometry. The pharmacokinetic parameters were calculated using WinNonlin 8.2 software, and the pharmacokinetic parameters were calculated using the non-compartmental model method.

[0457] 2. Experimental Results

[0458] The PK results of some compounds of the present invention on CD-1 mice are shown in Table 2 and Table 3.

[0459] Table 2 PK results of some compounds in the present invention in CD-1 mice (plasma)

[0460]

[0461] Table 3 PK results of some compounds in the present invention in CD-1 mice (lung)

[0462]

[0463] Conclusion: The compounds of the present invention are better orally absorbed in CD-1 mice than the reference compounds, and have better exposure in both plasma and lungs. Other compounds of the present invention have similar effects.

[0464] Example 18: Study on the antiviral activity of some compounds of the present invention in BALB / c mice

[0465] 1. Experimental Methods

[0466] Thirty 10-week-old female BALB / c mice were purchased from the Human SJA Experimental Animal Center (Changsha, China). The mice were housed in a specific pathogen-free environment under standard conditions. The mice in the prevention and treatment groups were orally administered with compound 25 or Lonafarnib 2 hours before or 2 hours after infection, respectively. The mice in the drug treatment group were orally administered with 20 / 40 mg kg -1 dose -1 (mpk) of compound or Lonafarnib, diluted in 200 μL 5% DMSO / 20% hydroxypropyl-β-cyclodextrin, twice a day (BID) for 4 consecutive days. Mice were anesthetized with isoflurane and inoculated intranasally with 1×10 6PFU of RSVA2. The body weight of mice was recorded every day. Mice were killed on the 4th day after infection, and lung tissue samples were taken for virological and histopathological analysis. After weighing the right lung tissue, 1000 μL PBS was added to the test tube, ground with a grinder, and 100 μL of ground tissue was taken to determine the virus titer by FFA method. Briefly, centrifugation was performed at 30°C (350g×30min), and 100 μL of ground tissue supernatant or 4-fold serial dilutions were used to infect cells. At 24hpi, cells were fixed with 4% PFA and then permeabilized with 0.3% Triton X-100. Anti-RSV-FITC (Cat. No. GTX36375) was used at a dilution of 1:100 in PBS and incubated overnight at 4°C. RSV was detected using anti-RSV-FITC (Cat. No. GTX36375) diluted 1:100 in PBS and incubated overnight at 4°C. Fluorescent spots were quantified using Image J. The virus titer was calculated according to the following formula: virus titer (FFU / g) = average number of fluorescent foci per well × dilution × volume index × volume × lung tissue weight index (converted to g). The left lung tissue was fixed with 4% formaldehyde, embedded, sliced, and stained with H&E to observe pathological changes. The sections were scored blindly by two independent pathologists according to the severity level of 0-4.

[0467] 2. Experimental Results

[0468] The in vivo efficacy of compound 25 and Lonafarnib against RSV infection in BALB / c mice is shown in Figure 1 Conclusion: At a dosage of 40 mpk, compound 25 has significantly improved anti-RSV activity in BALB / c mice compared with Lonafarnib.

[0469] Among them, positive control: Lonafarnib

[0470]

[0471] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the present invention itself. Various changes in form and details may be made without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. A tricyclic compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound has a structure of Formula I or Formula II: in, The tricyclic fragment ring I, ring II or ring III are each independently selected from C5-C 20 Aromatic ring, C5-C 20 Aromatic heterocycle, C5-C 20 Substituted aromatic ring or C5-C 20 Substituted aromatic heterocycle, the C5-C 20 The substituted aromatic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, and the C5-C 20 Substituted aromatic heterocycles are C5-C 20 The aromatic heterocyclic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN. 20 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S; Ring A is C5-C 15 Aliphatic ring, C5-C 15 Spirocyclic, C5-C 15 Heterospirocyclic, C5-C 15 Bridge ring, C5-C 15 Heterobridged ring, C5-C 15 Aliphatic heterocyclic, C5-C 15 Aromatic ring, substituted C5-C 15 Aliphatic ring or substituted C5-C 15 Aromatic ring, the C5-C 15 Heterospirocyclic, C5-C 15 Heterobridged ring or C5-C 15 The aliphatic heterocyclic ring contains 1 to 2 independent nitrogen atoms. 15 The aromatic heterocyclic ring contains 1 to 3 atoms independently selected from N or O, and the substituted C5-C 15 Aliphatic ring and C5-C 15 The aromatic rings are optionally substituted by 1 to 2 substituents independently selected from -CF3 or halogen; Ring B is C3-C 12 Aliphatic ring, C3-C 12 Spirocyclic, C3-C 12 Bridge ring, C3-C 12 Aliphatic heterocyclic, C5-C 12 Aromatic ring, C3-C 12 Aromatic heterocycle, substituted C3-C 12 Aliphatic ring, substituted C5-C 12 Aromatic ring or substituted C3-C 12 Aromatic heterocyclic ring, the C3-C 12 Aliphatic heterocyclic or C3-C 12 The aromatic heterocyclic ring contains 1 to 3 atoms independently selected from N or O, and the substituted C3-C 12 Aliphatic ring or substituted C5-C 12 Aromatic ring or C3-C 12 The aromatic heterocyclic ring is optionally substituted by 0 to 2 substituents independently selected from -CH3, -OCH3, -F, or -CF3. 12 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S; R 1 ,R 2 ,R 3 ,R 4 ,R 5 Each is independently selected from -H, -D, halogen, -CH3, -C2H5, -CF3, -SF5, -CD3, C3-C4 cycloalkyl, -OCH3, -OCF3, -NH2, -NO2 or -CN; X is -C-, -N or -NO-; Q is -C- or -N; Y, Z are each independently selected from -CH2-, -NH-, -N(CH3)-, -O-, -S-, -SO- or -SO2-; The chemical bond between Y and Z is a single bond or a double bond; The chemical bond between Q and A ring is a single bond or a double bond; T is -O-, -NH-, -CO-, -CHF-, -CF2- or -(CH2)n-, where n is any integer from 0 to 4; E is selected from -O-, -NH-, -CO-, -CHF-, -CF2-, -C(CH3)2-, Or -(CH2)n-, n = any integer from 0 to 4; L is selected from -O-, -NH-, -CO-, -SO-, -SO2-, or -(CH2) n -, n = any integer from 0 to 4; M is selected from -H, -D, -CH3, -CD3, -CF3, -OH, -NHR 6 ,-CONHR 6 、-SO2CH3、-CH2=NR 6 、-(CH2) n -, n = any integer from 0 to 4, C3-C6 aliphatic ring, C3-C6 aliphatic heterocyclic ring, C5-C 12 Aromatic ring or C5-C 12 Aromatic heterocycle, the C3-C6 aliphatic heterocycle or C5-C 12 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S; R 6 Selected from -H, -D, -OH or -OCH3.

2. The tricyclic compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The tricyclic fragment ring I, ring II or ring III are each independently selected from C5-C 15 Aromatic ring, C5-C 15 Aromatic heterocycle, C5-C 15 Substituted aromatic ring or C5-C 15 Substituted aromatic heterocycle, the C5-C 15 The substituted aromatic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, and the C5-C 15 Substituted aromatic heterocycles are C5-C 15 The aromatic heterocyclic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN. 15 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S; Ring A is C5-C 12 Aliphatic ring, C5-C 12 Spirocyclic, C5-C 12 Heterospirocyclic, C5-C 12 Bridge ring, C5-C 12 Heterobridged ring, C5-C 12 Aliphatic heterocyclic, C5-C 12 Aromatic ring, substituted C5-C 12 Aliphatic ring or substituted C5-C 12 Aromatic ring, the C5-C 12 Heterospirocyclic, C5-C 12 Heterobridged ring or C5-C 12 The aliphatic heterocyclic ring contains 1 to 2 independent nitrogen atoms. 12 The aromatic heterocyclic ring contains 1 to 3 atoms independently selected from N or O, and the substituted C5-C 12 Aliphatic ring and C5-C 12 The aromatic rings are optionally substituted by 1 to 2 substituents independently selected from -CF3 or halogen; Ring B is C4-C 11 Aliphatic ring, C4-C 11 Spirocyclic, C4-C 11 Bridge ring, C4-C 11 Aliphatic heterocyclic, C4-C 11 Aromatic ring, C4-C 11 Aromatic heterocycle, substituted C4-C 11 Aliphatic ring, substituted C4-C 11 Aromatic ring or substituted C4-C 11 Aromatic heterocyclic ring, the C4-C 11 Aliphatic heterocyclic or C4-C 11 The aromatic heterocyclic ring contains 1 to 3 atoms independently selected from N or O, and the substituted C4-C 11 Aliphatic ring or substituted C4-C 11 Aromatic ring or C4-C 11 The aromatic heterocyclic ring is optionally substituted by 0 to 2 substituents independently selected from -CH3, -OCH3, -F, or -CF3. 11 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S; L is selected from -O-, -NH-, -CO-, -SO2-, or -(CH2) n -, n = any integer from 0 to 4; M is selected from -H, -D, -CH3, -CD3, -CF3, -OH, -NHR 6 ,-CONHR 6 、-SO2CH3、-CH2=NR 6 、-(CH2) n -, n = any integer from 0 to 4, C3-C6 aliphatic ring, C3-C6 aliphatic heterocyclic ring, C5-C 10 Aromatic ring or C5-C 10 Aromatic heterocycle, the C3-C6 aliphatic heterocycle or C5-C 10 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S.

3. The tricyclic compound or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The tricyclic fragment ring I, ring II or ring III are each independently selected from C5-C 10 Aromatic ring, C5-C 10 Aromatic heterocycle, C5-C 10 Substituted aromatic ring or C5-C 10 Substituted aromatic heterocycle, the C5-C 10 The substituted aromatic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN, and the C5-C 10 Substituted aromatic heterocycles are C5-C 15 The aromatic heterocyclic rings are optionally substituted by 0 to 3 substituents independently selected from -CF3, halogen or -CN. 10 The aromatic heterocyclic ring contains 1 to 3 heteroatoms independently selected from N, O or S; Ring A is C5-C 10 Aliphatic ring, C5-C 10 Aliphatic heterocyclic, substituted C5-C 10 Aliphatic ring or substituted C5-C 10 Aliphatic heterocyclic ring, the C5-C 10 Aliphatic heterocyclic or substituted C5-C 10 The aliphatic heterocyclic ring contains 1 to 2 independent nitrogen atoms, and the substituted C5-C 10 Aliphatic ring and C5-C 10 The aliphatic heterocycle is optionally substituted by 1 to 2 substituents independently selected from -CF3 or halogen; Ring B is a C4-C6 aliphatic heterocycle, C6-C 10 Spirocyclic ring, C7-C8 bridged ring, C4-C8 aromatic ring, substituted C4-C6 aliphatic ring or substituted C4-C8 aromatic ring, the C4-C6 aliphatic heterocyclic ring contains 1 to 2 independently selected N atoms, the substituted C4-C 11 The aromatic rings are each optionally substituted with 0 to 2 substituents independently selected from -CH3, -OCH3, -F, or -CF3; L is selected from -NH-, -CO-, -SO2- or -(CH2) n -, n = any integer from 0 to 4; M is selected from -CH3, -CD3, -OH, -NHR 6 ,-CONHR 6 、-SO2CH3、-CH2=NR 6 、-(CH2) n -, n=any integer from 0 to 4, a C3-C6 aliphatic ring or a C5-C6 aromatic ring, wherein the C5-C6 aromatic heterocyclic ring contains 1 to 2 independent N atoms.

4. The tricyclic compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The tricyclic fragment ring I, ring II or ring III are each independently selected from any one of the following groups:

5. The tricyclic compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The ring A is selected from any one of the following groups:

6. The tricyclic compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The ring B is selected from any one of the following groups:

7. The tricyclic compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The R 1 ,R 2 ,R 3 are each independently selected from -H, -D, -F, -Cl or -Br; R 4 ,R 5 Each is independently selected from -H, -D, -F, -CH3, -CF3, -OCH3 or -OCF3.

8. The tricyclic compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The structure of the -LM is as shown in Formula III or Formula IV: Wherein V is selected from -O-, -NH-, -CH2-, -CO-, -SO- or -SO2-, and n = any integer from 0 to 4; U is selected from -CH-, -CH2-, or n = any integer between 0 and 4; R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3, -CF3-, -OCH3, -OCF3, -COOH, -CONH2, -SO2NH2, -SO2CH3, -isopropyl, -cyclopropyl, -cyclohexyl or The R 9 Selected from -H, -OH, -NH2, -OCH3, -CH3, -CD3, -CF3-, -OCF3 or halogen; R 8 Selected from -H, -CH3 or -CD3.

9. The tricyclic compound or a pharmaceutically acceptable salt thereof according to claim 8, characterized in that: The structure of the -LM is as shown in Formula III or Formula IV: Wherein V is selected from -CH2-, -CO-, -SO- or -SO2-, and n = any integer from 0 to 3; U is selected from -CH-, -CH2-, n = any integer between 0 and 3; R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3, -CONH2, -isopropyl, -cyclopropyl, -cyclohexyl or The R 9 Selected from -H, -OH, -NH2, -OCH3, -CH3, -CD3, -CF3-, -OCF3 or halogen; R 8 Selected from -H or -CH3.

10. The tricyclic compound or pharmaceutically acceptable salt thereof according to claim 9, characterized in that: The structure of the -LM is as shown in Formula III or Formula IV: Wherein V is selected from -CH2-, -CO-, and n = any integer from 1 to 2; U is selected from -CH-, -CH2-, n = any integer between 0 and 2; R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3, -CONH2, -isopropyl, -cyclopropyl or -cyclohexyl; R 8 Selected from -H or -CH3.

11. The tricyclic compound or a pharmaceutically acceptable salt thereof according to claim 10, characterized in that: The structure of the -LM is as shown in Formula III or Formula IV: wherein V is selected from -CO-, n=1; U is selected from -CH- or -CH2-, n = any integer from 0 to 2; R 7 Selected from -H, -OH, -NH2, -N-OH, -N-OCH3, -CH3, -CD3 or -CONH2; R 8 Selected from -H or -CH3.

12. The tricyclic compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: Select any of the following compounds:

13. A pharmaceutical composition, characterized in that Comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate thereof and a pharmaceutically acceptable carrier.

14. The pharmaceutical composition according to claim 13, characterized in that The pharmaceutically acceptable salt is a salt of the compound formed with the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, carbonic acid, nitric acid, hydrobromic acid, hydroiodic acid, maleic acid, fumaric acid, tartaric acid, citric acid, malic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, succinic acid, acetic acid, mandelic acid, isobutyric acid or malonic acid.

15. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 13 or 14, in the preparation of a medicament for preventing or treating viral infection.

16. The use according to claim 15, characterized in that The viral infection is preferably a respiratory syncytial virus infection.

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