Protease inhibitor for treating or preventing virus infection and application thereof

Abstract

The invention belongs to the field of anti-infection drugs, and discloses a protease inhibitor for treating or preventing virus infection and application thereof. The protease inhibitor is a compound represented by the following formula (I) and pharmaceutically acceptable salts, esters, prodrugs, solvates and isomers thereof: the compound is a novel protease inhibitor, can inhibit proteins encoded by coronavirus or interfere with the life cycle of the coronavirus, and can also be used as an antiviral agent, such as an anti-inflammatory agent, an anti-inflammatory agent and an anti-inflammatory agent. The 3CL protease inhibitor is used for treating respiratory system diseases, and compared with the prior art, the 3CL protease inhibitor provided by the invention is high in activity, lower in cytotoxicity, high in metabolic stability in liver microsomes, higher in bioavailability, better in drug absorption, distribution, metabolism and excretion properties, free of inhibition on main CYP enzymes and lower in DDI risk.

Description

Technical Field

[0001] This invention relates to the field of anti-infective drugs, and more specifically to a protease inhibitor for treating or preventing viral infections and its uses. Background Technology

[0002] The novel coronavirus (Coronavirus Disease 2019, COVID-19), commonly known as "COVID-19" by the World Health Organization, is primarily transmitted through direct contact, aerosol transmission, and contact. Since the end of 2019, people around the world have been infected with the novel coronavirus, accelerating research into the virus and the development of related drugs. Drug development and approval processes have sped up, and an increasing number of "COVID-19 specific drugs" have been released through special approval processes.

[0003] The mechanisms of action of "COVID-19 specific drugs" mainly involve RdRp inhibitors and 3CL protease inhibitors. RdRp inhibitors work by targeting the RdRp site of the novel coronavirus to precisely block its RNA replication; examples include Remindevir, Molnupiravir, Azvudine, and Remdesivir. 3CL protease inhibitors work by inhibiting viral protein cleavage, thereby blocking viral replication; examples include Ensitrelvir Fumaric Acid (XOCOVA), Paxlovid, Cilgavimab / tixagevimab (Evusheld), and SIM-0417 (Senotvir / ritonavir).

[0004] Although many drugs have been approved, many entered the market through special approval processes in various countries, such as emergency use authorizations in Japan and South Korea, conditional approvals in China, and orphan drug and fast track approvals in the United States. On the one hand, to date, M... pro The types and structures of inhibitors are relatively limited. In particular, existing covalent main protease inhibitors often use highly reactive structures such as aldehydes and α,β-unsaturated carbonyl groups as covalent warheads, posing potential drug safety risks. On the other hand, non-covalent inhibitors generally exhibit low inhibitory activity against main proteases. Therefore, discovering highly active and selective SARS-CoV-2 main protease inhibitors is an important direction for current anti-COVID-19 drug development. Furthermore, pneumonia caused by the novel coronavirus remains a significant public health problem now and in the future. 3CL protease inhibitors for the treatment or prevention of coronavirus infection are an important supplement to vaccines for the current COVID-19 pandemic and future coronavirus outbreaks. Summary of the Invention

[0005] In response to the current COVID-19 pandemic and the potential crisis of future coronavirus outbreaks, this invention provides a protease inhibitor for the treatment or prevention of viral infections and its uses. The protease inhibitor provided by this invention can inhibit proteins encoded by coronaviruses or interfere with the life cycle of coronaviruses, and can also be used as an antiviral agent against small RNA viruses. Compared with existing technologies, the 3CL protease inhibitor provided by this invention has high activity, lower cytotoxicity, high metabolic stability in liver microsomes, higher bioavailability, and superior absorption, distribution, metabolism, and excretion properties. Furthermore, it does not inhibit major CYP enzymes and has a lower risk of drug-induced drug dispersive disorder (DDI).

[0006] To achieve the objectives of this invention, the present invention provides compounds of formula (I) and their pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof:

[0007]

[0008] in:

[0009] The R1 is either absent or contains a deuterium or halogen atom, or is a C1-C4 alkyl group, or a C3-C6 cycloalkyl group, or a -C0-C4 alkylsilyl group;

[0010] The R2 is selected from hydrogen, deuterium, C1-C4 alkyl, -(C0-C4 alkyl)-(C3-C6 cycloalkyl), -C0-C4 alkylsilyl, and -(C0-C4 alkyl)-phenyl; the C1-C4 alkyl, -C0-C4 alkylsilyl, and -(C0-C4 alkyl)-phenyl have one or more substituents selected from alkyl, amino, oxygen, alkoxy, hydroxyl, carboxyl, and halogen atoms;

[0011] The R3 is selected from the halogenated atom, C3-C6 cycloalkyl, hydroxyl, amino, C1-C6 alkylamine, C1-C6 alkoxy, C1-C 18 Acyloxy group, C1-C 18 amide group, C1-C 18 A C1-C6 alkyl group substituted with one or more substituents in the oxycarbonyl group, wherein the substituents are optionally further replaced by an amino group, a hydroxyl group, a carboxyl group, or a (C1-C6 alkyl) group. m Amine group, (C1-C6 alkyl group) m Ammonium, C6-C 20 aryl-substituted C1-C6 alkoxy, C6-C 20 Heteroaryl-substituted C1-C6 alkoxy groups, -(OC2H4) n One or more of O (C1-C6 alkyl) are substituted;

[0012] Where m is selected from 1 to 3, and n is selected from 1 to 10;

[0013] The R4 is selected from NHR5, -CHR2NHSO2-(C1-C3 alkyl), -CHR2NHSO2-(C1-C3 haloalkyl), -CHR2NHC(=O)-O-(C1-C3 alkyl), OR5, or R5, wherein R5 is selected from the following groups, and R5 is optionally substituted:

[0014]

[0015]

[0016] Furthermore, in some embodiments of the present invention, R1 is absent or contains deuterium or halogen atoms.

[0017] Furthermore, in some embodiments of the present invention, R1 is absent.

[0018] Furthermore, in some embodiments of the present invention, R2 is selected from C1-C4 alkyl, -(C0-C4 alkyl)Si(CH3)3, -(C0-C4 alkyl)-phenyl, -(C0-C4 alkyl)-(C3-C6 cycloalkyl).

[0019] Furthermore, in some embodiments of the present invention, R2 is selected from C1-C4 alkyl, -(C1-C2 alkyl)Si(CH3)3, and -(C1-C3 alkyl)-cyclopropyl.

[0020] Furthermore, in some embodiments of the present invention, R3 is selected from the following substituents:

[0021]

[0022]

[0023] Furthermore, in some embodiments of the present invention, R4 is selected from...

[0024] In some specific embodiments of the present invention, the present invention provides the following compounds and their pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers:

[0025]

[0026]

[0027]

[0028]

[0029] Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. Additional aspects and advantages of this invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. It should be understood that the following description is merely illustrative and not intended to limit the invention.

[0031] definition

[0032] The following lists the definitions of the terms used to describe the invention. Unless otherwise limited in specific circumstances, these definitions apply when a term is used alone or as part of a larger group in this specification and claims.

[0033] As used herein, the term "substituted" means that one, two, three, or more hydrogen atoms thereon are independently replaced by substituents including, but not limited to, -F, -Cl, -Br, -I, -OH, C. 1- C 12 -alkyl; C2-C 12 -Alkenyl, C2-C 12 -alkynyl group, -C3-C 12 -cycloalkyl, protected hydroxyl, -NO2, -N3, -CN, -NH2, protected amino, oxygen, thio, -NHC 1- C 12 -alkyl, -NH-C2-C8-alkenyl, -NH-C2-C8-ynyl, -NH-C3-C 12 -cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino, -OC 1- C 12 -alkyl, -O-C2-C8-alkenyl, -O-C2-C8-ynyl, -O-C3-C 12 -cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)-C 1- C 12 -alkyl, -C(O)-C2-C8-alkenyl, -C(O)-C2-C8-ynyl, -C(O)-C3-C 12 -cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocycloalkyl, -CONH2, -CONH-C 1- C 12 -alkyl, -CONH-C2-C8-alkenyl, -CONH-C2-C8-ynyl, -CONH-C3-C12 -cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -OCO2-C 1- C 12 -alkyl, -OCO2-C2-C8-alkenyl, -OCO2-C2-C8-ynyl, -OCO2-C3-C 12 -cycloalkyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2-heterocycloalkyl, -CO2-C 1- C 12 Alkyl, -CO2-C2-C8 alkenyl, -CO2-C2-C8 ynyl, CO2-C3-C 12 -cycloalkyl, -CO2-aryl, CO2-heteroaryl, CO2-heterocycloalkyl, -OCONH2, -OCONH-C1C 12 -alkyl, -OCONH-C2-C8-alkenyl, -OCONHC2-C8-ynyl, -OCONH-C3-C 12 -cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocyclic-alkyl, -NHC(O)H, -NHC(O)-C 1- C 12 -alkyl, -NHC(O)-C2-C8-alkenyl, -NHC(O)-C2-C8-ynyl, -NHC(O)-C3-C 12 -cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocyclic-alkyl, -NHCO2-C 1- C 12 -alkyl, -NHCO2-C2-C8-alkenyl, -NHCO2-C2-C8-ynyl, -NHCO2-C3-C 12 -cycloalkyl, -NHCO2-aryl, -NHCO2-heteroaryl, -NHCO2-heterocycloalkyl, -NHC(O)NH2, -NHC(O)NH-C 1- C 12 -alkyl, -NHC(O)NH-C2-C8-alkenyl, -NHC(O)NHC2-C8-ynyl, -NHC(O)NH-C3-C 12 -cycloalkyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocycloalkyl, NHC(S)NH2, -NHC(S)NH-C 1- C 12 -alkyl, -NHC(S)NH-C2-C8-alkenyl, -NHC(S)NH-C2-C8-ynyl, -NHC(S)NH-C3-C 12-cycloalkyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH2, -NHC(NH)NH-C 1- C 12 -alkyl, -NHC(NH)NH-C2-C8-alkenyl, -NHC(NH)NH-C2-C8-alkynyl, -NHC(NH)NH-C3-C 12 -cycloalkyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocycloalkyl, -NHC(NH)-C 1- C 12 -alkyl, -NHC(NH)-C2-C8-alkenyl, -NHC(NH)-C2-C8-ynyl, -NHC(NH)-C3-C 12 -cycloalkyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocycloalkyl, -C(NH)NH-C 1- C 12 -alkyl, -C(NH)NH-C2-C8-alkenyl, -C(NH)NH-C2-C8-ynyl, -C(NH)NH-C3-C 12 -cycloalkyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocycloalkyl, -S(O)-C 1- C 12 -alkyl, -S(O)-C2-C8-alkenyl, -S(O)-C2-C8-ynyl, -S(O)-C3-C 12 -cycloalkyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocycloalkyl, -SO2NH2, -SO2NH-C 1- C 12 -alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-ynyl, -SO2NH-C3-C 12 -cycloalkyl, -SO2NH-aryl, -SO2NH-heteroaryl, -SO2NH-heterocycloalkyl, -NHSO2-C 1- C 12 -alkyl, -NHSO2-C2-C8-alkenyl, -NHSO2-C2-C8-ynyl, -NHSO2-C3-C 12 -cycloalkyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocycloalkyl, -CH2NH2, -CH2SO2CH3, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -C3-C 12-cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, -SC 1- C 12 -alkyl, -S-C2-C8-alkenyl, -S-C2-C8-ynyl, -S-C3-C 12 -cycloalkyl, -S-aryl, -S-heteroaryl, -S-heterocycloalkyl, or methylthio-methyl. In some embodiments, the substituents are independently selected from halogens, preferably Cl and F; C 1- C4-alkyl, preferably methyl and ethyl; halogenated-C 1- C4-alkyl, such as fluoromethyl, difluoromethyl, and trifluoromethyl; C2-C4-alkenyl; halo-C2-C4-alkenyl; C3-C6-cycloalkyl, such as cyclopropyl; C 1- C4-alkoxy groups, such as methoxy and ethoxy groups; halogenated-C 1- C4-alkoxy group, such as fluoromethoxy, difluoromethoxy, and trifluoromethoxy; acyl group; -CN; -OH; NH2; C 1- C4-alkylamino; di(C 1- C4-alkyl)amino; and NO2. It should be understood that aryl, heteroaryl, alkyl, etc., can be further substituted. In some cases, each substituent of the substituted portion is optionally additionally substituted by one or more groups, each group independently selected from C4-alkyl. 1- C4-alkyl; -CF3, -OCH3, -OCF3, -F, -Cl, -Br, -I, -OH, -NO2, -CN, and -NH2. Preferably, the substituted alkyl group may be further substituted with one or more halogen atoms, more preferably, with one or more fluorine or chlorine atoms.

[0034] The compounds described herein contain one or more centers of symmetry, and thus form enantiomers, diastereomers, and other stereoisomers. This invention is intended to include all such possible isomers, as well as their racemic and optically pure forms.

[0035] Antiviral activity

[0036] In some embodiments, the present invention provides a method for treating or preventing viral infections in a host in need, comprising administering a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt / ester thereof to the host. The viral infection is preferably a coronavirus infection. In some embodiments, the coronavirus is SARS-CoV-1, SARS-CoV-2, or MERS-CoV. Preferably, the coronavirus is SARS-CoV-2.

[0037] Synthesis method

[0038] The following examples illustrate the methods for preparing the compounds of this invention, which will provide a better understanding of the invention. These examples are intended to be illustrative only and are not intended to limit the scope of the invention. The synthetic methods for compounds not described in detail in this invention are similar to the specific compounds synthesized in the following examples, and can be obtained by referring to the examples below.

[0039] In the embodiments of this invention, 1A / a, 2A / a, 3A / a, 1B / b, 2B / b, etc., are only for the convenience of referring to a certain compound in the specific synthetic process route. Unless otherwise specified, the compounds referred to by 1A, 1B, etc. in different embodiments are not necessarily the same or different.

[0040] Example 1

[0041] The synthetic route for compound C10-280 is shown below:

[0042]

[0043] 1. Synthetic intermediate 2A

[0044] Cb2Cl was added to a DCM solution of 1A and DIPEA. The reaction mixture was stirred at 20°C for 5 h. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under vacuum. The residue was purified by rapid column chromatography (Combi-flash, 10 g silica gel, PE / EtOAc 0–10% gradient) to give 2A. LC-MS (ESI+): m / z = 374.2 [M+Na] + .

[0045] 2. Synthetic intermediate 3A

[0046] The solution of 2A in DCM / TFA (1:1, 10 mL) was stirred at 20 °C for 1 h. The mixture was then concentrated under vacuum to obtain 3A. 1 HNMR (400MHz, CDCl3) δ10.07(s,1H),7.19~7.28(m,5H),5.06-5.11(m,2H),4.20-4.37(m,1Hz),1.12-1.17(m,1H),0.90-0.96(m,1H),0.03(s,9H).

[0047] 3. Synthetic intermediate 4A

[0048] T3P was added to the DCM solutions of 3A, 3B, and DIPEA, and the mixture was stirred at 25°C for 2 h. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under vacuum. The residue was purified by rapid column chromatography to give 4A. LC-MS (ESI+): m / z = 509.3 [M+H] + .

[0049] 4. Synthetic intermediate 5A

[0050] A solution of 4A, TsOH-H2O, and Pd / C in MeOH was stirred at 25°C for 1 h under H2 (1 atm). The reaction mixture was filtered through a diatomaceous earth mat, and the filtrate was concentrated under vacuum to obtain 5A. LC-MS (ESI+): m / z = 375.2 [M+H] + .

[0051] 5. Synthetic intermediate 6A

[0052] Dissolve 5A and 5B in MeOH and stir at 0°C for 10 minutes, then add sodium cyanoborohydride. Stir the mixture at 25°C for 2 hours. Pour the reaction mixture into water and extract with EtOAc. Wash the combined organic layers with brine, dry with Na₂SO₄, and concentrate under vacuum. Purify the residue by rapid column chromatography to give 6A. LC-MS (ESI+): m / z = 640.3 [M+H] + .

[0053] 6. Synthetic intermediate 6B

[0054] DIPEA and DPPA were added to a toluene solution of 4,4-difluorocyclohexane-1-carboxylic acid at 25°C. After stirring at 25°C for 1 h, the reaction mixture was heated to 110°C and maintained for 13 h. The reaction was then cooled to 25°C to give a toluene solution of 6B.

[0055] 7. Synthetic intermediate 7A

[0056] Add 6B (30 mL toluene solution) to a solution of 6A in toluene (30 mL), and stir the reaction mixture at 60 °C for 2 h. Dilute the mixture with H₂O and extract with EtOAc (100 mL * 2). Wash the organic layer with brine, dry with Na₂SO₄, and concentrate under vacuum. Purify the residue by rapid chromatography to give 7A.

[0057] 8. Synthetic intermediate 8A

[0058] Piperidine was added to the DCM solution of 7A, and the reaction was stirred at 25°C for 1 hour. The reaction mixture was concentrated to give 8A, which was used directly in the next reaction without any purification.

[0059] 9. Synthetic intermediate 9A

[0060] 8A was added to a DMF solution of 8B, HATU, and DIPEA, and the reaction mixture was stirred at 25°C for 2 h. The reaction mixture was then quenched with water and extracted with EtOAc (20 mL * 2). The combined organic layers were washed with brine, dried over Na₂SO₄, and concentrated under vacuum. The residue was purified by rapid column chromatography to give 9A. LC-MS (ESI+): m / z = 922.2 [M + Na] + .

[0061] 10. Synthetic intermediate 10A

[0062] Burgess reagent was added to a THF solution of 9A, and the mixture was incubated at 25°C for 2 hours. The mixture was diluted with H₂O and extracted with EtOAc. The organic layer was washed with brine, dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography to give 10A. LC-MS (ESI+): m / z = 882.2 [M+H] + .

[0063] 11. Synthesized product 11A (C10-280)

[0064] Piperidine was added to a DCM solution of 10A, and the reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was then concentrated under vacuum. The residue was purified by rapid column chromatography to give 11A. LC-MS (ESI+): m / z = 660.2 [M+H] + . 1 H NMR (400MHz, DMSO) δ10.76(s,1H),8.36(s,1H),7.24(t,J=7.6Hz,1H),6.89(dt,J=15.4,6.7 Hz,4H),5.23–5.15(m,2H),4.11(d,J=10.6Hz,1H),3.73(d,J=10.6Hz,1H),3.41(s,1H),3.1 8–3.12(m,4H),3.03(d,J=5.0Hz,1H),2.68–2.62(m,1H),2.51–2.47(m,1H),2.07–1.84(m,4 H),1.75–1.67(m,2H),1.56–1.49(m,1H),1.25–1.11(m,2H),0.95–0.75(m,8H),0.00(s,9H).

[0065] Example 2

[0066] The synthetic route for compound C10-284 is shown below:

[0067]

[0068] 1. Synthetic intermediate 2A

[0069] Burgess reagent was added to a THF solution of 1A, and the reaction mixture was stirred at 25°C for 2 h. The mixture was diluted with H₂O and extracted with EtOAc. The organic layer was washed with brine, dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography to give 2A. LC-MS (ESI+): m / z = 805.2 [M + Na] + . 1 H NMR (400MHz, DMSO) δ10.74(s,1H),7.93(d,J=7.5Hz,2H),7.72(d,J=7.4Hz,2H),7.58–7.54(m,1H),7.46(t,J=6.6Hz,2H),7 .37(td,J=7.4,1.4Hz,2H),7.23(t,J=7.7Hz,1H),6.97–6.85(m,3H),6.21(d,J=7.3Hz,1H),5.17(dt,J=27.2,7.8Hz,2H),4. 46(d,J=6.4Hz,2H),4.28(t,J=6.3Hz,1H),4.06(d,J=10.6Hz,1H),3.75(d,J=10.6Hz,1H),3.43–3.37(m,1H),3.19–3.00(m, 4H),2.70–2.62(m,1H),2.50–2.45(m,1H),2.00–1.66(m,6H),1.13–1.12(m,3H),0.77(dd,J=14.2,6.5Hz,1H),0.00(s,9H).

[0070] 2. Synthetic intermediate 3A

[0071] Piperidine was added to the DCM solution of 2A, and the reaction was stirred at 25°C for 1 h. The reactants were concentrated under vacuum, and the residue was purified by rapid column chromatography (25 g silica gel, DCM / MeOH 0–10% gradient) to obtain 3A. LC-MS (ESI+): m / z = 561 [M+H] + . 1H NMR(400MHz,DMSO)δ7.65(d,J=6.0Hz,1H),7.27–7.21(m,1H),7.01–6.96(m,2H),6 .90(d,J=7.7Hz,1H),5.20–5.10(m,2H),4.05(d,J=10.6Hz,1H),3.72(d,J=10.7Hz, 1H),3.14(t,J=5.7Hz,2H),2.73–2.58(m,3H),2.51–2.44(m,1H),2.01–1.57(m,6H) ,1.48–1.41(m,1H),1.28–1.09(m,3H),0.79(dd,J=14.4,6.4Hz,1H),-0.00(s,9H).

[0072] 3. Synthetic product 4A (C10-284)

[0073] 3A was added to a DMF mixture of 3B, EDCI, HOBt, and DIPEA, and the reaction was stirred at 25°C for 2 h. The reaction mixture was then quenched with water and extracted with EtOAc (10 mL * 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under vacuum. The residue was purified by rapid column chromatography (25 g silica gel, DCM / MeOH 0–10% gradient) to obtain the crude product. The crude product was purified by Prep-HPLC (Waters MS triggered Prep-HPLC with Acquity QDA detector (Welch 10 u C18 250 x 21.2 mm, 20 mL / min, MeCN / H2O + 0.1% FA: 55%–65%)) to obtain C10-284. LC-MS (ESI+): m / z = 853.3 [M + H) + . 1H NMR (400MHz, DMSO) δ10.74(s,1H),8.20(t,J=5.6Hz,1H),7.27–7.22(m,1H),6.95–6.89(m,3H),6.64(d,J=7.2Hz,1H), 5.21(t,J=7.6Hz,1H),5.11(t,J=7.9Hz,1H),4.08(d,J=10.7Hz,1H),3.99(s,2H),3.73(d,J=10.6Hz,1H),3.63–3.61( m,4H),3.58–3.52(m,12H),3.45–3.44(m,2H),3.40–3.35(m,3H),3.26(s,3H),3.20–3.10(m,4H),2.70–2.64(m,1H),2 .50–2.46(m,1H),2.02–1.62(m,6H),1.54–1.46(m,1H),1.29–1.17(m,2H),0.78(dd,J=14.2,6.6Hz,1H),0.00(s,9H).

[0074] Example 3

[0075] The synthetic route for compound C10-390 is shown below:

[0076]

[0077] 1. Synthesize compound 2A

[0078] Dissolve 1A and 1B in dichloroethane, add sodium triacetylborohydride, and react at room temperature for 16 hours after the addition is complete. Quench the reaction with saturated sodium bicarbonate solution (50 mL), extract the reaction solution with dichloromethane (30 mL × 3), dry to anhydrous sodium sulfate, filter, and concentrate to obtain 2A. LC-MS (ESI+): m / z = 322 [M+1] + .

[0079] 2. Synthesis of compound 3A

[0080] 2A, 2B, and DIPEA were added to DMF and stirred until dissolved. A 50% T4P / DMF solution was added at room temperature, and the mixture was stirred for 2 hours at room temperature. The reaction solution was quenched with water, extracted twice with DCM, and the combined organic phases were washed with 5% citric acid aqueous solution and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 3A. LC-MS (ESI+): m / z = 463 [M-55] + .

[0081] 3. Synthesize compound 4A

[0082] 3A was dissolved in DCM (16 mL) and TFA (16 mL) with stirring and reacted at room temperature for 1 h. The reaction solution was concentrated at 35 °C to obtain 4A. LC-MS (ESI+): m / z = 463 [M+H] + .

[0083] 4. Synthesis of compound 5A

[0084] 4A was dissolved in DMF, HOBT was added at 0°C, and the mixture was stirred for 10 minutes. Then, EDCI and 4B were added, and the reaction was allowed to proceed at room temperature for 2 hours. The reaction mixture was quenched with water, extracted twice with DCM, and the combined organic phases were washed with 5% citric acid aqueous solution and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI-): m / z = 674 [M-1] - .

[0085] 5. Synthesize compound 6A

[0086] 5A was dissolved in methanol, and 10% palladium on carbon (80 mg) and 20% palladium hydroxide (80 mg) were added. The mixture was purged three times with hydrogen and hydrogenated at room temperature for 16 h. The reaction solution was filtered, concentrated, and purified by column chromatography to obtain 6A. LC-MS (ESI-): m / z = 584 [M⁻¹] - .

[0087] 6. Synthesize compound 7A

[0088] 6A was dissolved in dichloromethane and tetrahydrofuran, and N,N-diisopropylethylamine was added. Acetic anhydride was added at 0°C, and the reaction was allowed to proceed at room temperature for 1 hour after the addition was complete. The reaction was quenched with saturated ammonium chloride aqueous solution, and the pH was adjusted to 5 with 10% citric acid aqueous solution. The mixture was extracted twice with dichloromethane and dried. The solution was concentrated and purified by column chromatography to obtain 7A. LC-MS (ESI-): m / z = 626 [M⁻¹] - .

[0089] 7. Synthesize compound 8A (C10-426)

[0090] 7A was dissolved in ultradry dichloromethane (5 mL) and ultradry tetrahydrofuran (5 mL), and Burgess reagent was added at room temperature. The reaction mixture was reacted for 0.5 hours, quenched with water (100 mL) and dichloromethane (200 mL), and the layers were separated. The organic phase was washed with 5% sodium chloride aqueous solution, dried, and purified by column chromatography to obtain C10-426. LC-MS (ESI-): m / z = 608 [MH] - . 1H-NMR (400MHz, DMSO-d6)δ=12.74(d,1H),10.68(d,1H),7.23-7.08(m,4H),6.97-6.84(m,2H),5.22 (dt,1H),4.08(s,2H),3.86-3.78(m,2H),1.87(s,4H),1.57(s,2H),1.24(s,2H),1.01-0.85(m,9H).

[0091] 8. Synthesized product 9A (C10-390)

[0092] 8A was dissolved in methanol (40 mL), and 13% potassium carbonate aqueous solution (8 mL) was added. The reaction was carried out at room temperature for 0.5 h. The reaction was quenched with saturated ammonium chloride aqueous solution (50 mL), and the pH was adjusted to 7 with 10% citric acid aqueous solution. Extraction was performed twice with dichloromethane (100 mL), and the sample was dried. The solution was concentrated. Purification by column chromatography yielded C10-390. LC-MS (ESI-): m / z = 566 [M⁻¹] - . 1 H-NMR (400MHz, DMSO-d6)δ=7.19-6.68(m,6H),5.29(s,2H),3.92-3.77(m,2H),2. 74-2.63(m,2H),1.84(d,2H),1.65(s,2H),1.55(t,1H),1.25(d,2H),0.95(s,6H).

[0093] Example 4

[0094] The synthetic route for compound C10-392 is shown below:

[0095]

[0096] 1. Synthetic intermediate 2A

[0097] 1A, 1B, dichloromethane, N,N-diisopropylethylamine, and 4-dimethylaminopyridine were reacted at room temperature for 1 hour. LCMS showed that the reaction was complete. The reaction solution was washed with purified water, and the organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated to obtain the crude product, which was purified by column chromatography to obtain 2A. LCMS (ESI-): m / z = 876.7 [M-1] - .

[0098] 2. Synthetic product 3A (C10-392)

[0099] Add 2A, dichloromethane, and Burgess reagent to a three-necked flask and stir at room temperature for 2 hours. Purify by column chromatography to obtain C10-392. LCMS (ESI-): m / z = 585.7 [M⁻¹] - .1 H NMR(400MHz,Chloroform-d)δ9.78(s,1H),8.43(s,1H),7.18(m,1H),6.96– 6.86(m,3H),6.76-6.61(m,2H),5.34(m,1H),5.02(m,1H),4.38(m,2H),4.23 (m,2H),4.06(m,2H),3.63–3.57(m,16H),3.34(m,3H),2.82(m,1H),2.55(m, 1H),1.95(m,1H),1.61(m,1H),1.23(m,2H),1.01(m,1H),1.02–0.77(m,6H).

[0100] Example 5

[0101] The synthetic route for compound C10-440 is shown below:

[0102]

[0103] 1. Synthetic intermediate 2A

[0104] 1A, DCM, and DIEA were added to a three-necked flask, and the mixture was cooled to 0°C. Benzyl chloroformate was slowly added dropwise, and the mixture was stirred at room temperature for 2 hours. LC-MS analysis showed that the product comprised 80% of the product. The reaction was stopped, the reaction solution was washed with water, and the mixture was separated. The organic phase was dried over anhydrous sodium sulfate, the filtrate was concentrated, and purified by column chromatography to obtain 2A. LC-MS (ESI+): m / z = 222.2 [M-BOC+1] + .

[0105] 2. Synthetic intermediate 3A

[0106] Add 2A, DMF, and sodium hydride to a three-necked flask, stir at room temperature for 30 min, then add 2B, stir at room temperature for 1 hour. LCMS analysis showed that the starting materials had reacted completely, and the product was the main peak. Stop the reaction, add water (40 mL), extract with ethyl acetate (20 mL * 2), wash the organic phase with saturated brine, and dry with anhydrous sodium sulfate. Concentrate the filtrate to obtain the crude product, and purify by column chromatography to obtain 3A. LCMS (ESI+): m / z = 268.0 [M – BOC+1] + .

[0107] 3. Synthetic intermediate 4A

[0108] Add 3A and methanol to a three-necked flask, add hydrochloric acid dropwise, then add palladium on carbon, and purge with hydrogen three times. Stir at room temperature for 2 hours. LCMS monitoring showed that the reactants had reacted completely. Stop the reaction, filter the reaction mixture with diatomaceous earth, and concentrate the filtrate to obtain 4A. LCMS (ESI+): m / z = 268.0 [M+1] +.

[0109] 4. Synthetic intermediate 5A

[0110] Add 4A, 4B, DMF, DIEA, and HATU to a three-necked flask, heat to 90℃ and stir for 4 hours. LCMS analysis showed that the starting material reaction was complete and the product percentage was 90%. Stop the reaction, add water (20 mL), and extract with ethyl acetate (10 mL * 2). Wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate to obtain the crude product, and purify by column chromatography to obtain 5A. LCMS (ESI-): m / z = 429.2 [M-1] - .

[0111] 5. Synthetic intermediate 6A

[0112] 5A, trifluoroacetic acid, and DCM were added to a three-necked flask and stirred at room temperature for 30 minutes. LCMS analysis showed that the reactants had reacted completely. The reaction was stopped, and the reaction solution was concentrated to obtain 6A. LCMS (ESI-): m / z = 373.1 [M-1] - .

[0113] 6. Synthetic intermediate 7A

[0114] Add 6A and DMF to a three-necked flask, cool to 0°C, add HOBT, stir for 10 min, then add 6B and EDCI, and return to room temperature and stir for 1 hour. LCMS analysis showed that the starting material reaction was complete. Stop the reaction, quench the reaction solution with water (10 mL), extract with ethyl acetate (10 mL * 2), wash the organic phase with saturated citric acid aqueous solution, saturated sodium bicarbonate, and saturated brine, dry with anhydrous sodium sulfate, concentrate to obtain the crude product, and purify by thin-layer chromatography to obtain product 7A. LCMS (ESI-): m / z = 586.3 [M-1] - .

[0115] 7. Synthesized product 8A (C10-440)

[0116] 7A, Burgess reagent, and DCM were added to a single-sided plate, and the mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was purified by thin-layer chromatography to obtain C10-440. LCMS (ESI-): m / z = 568.3 [M-1] - . 1H NMR(400MHz,DMSO-d6)δ12.66(s,1H),10.62(s,1H),7.26-7.03(m,3H),7.00-6.79(m,2H),6.63(d,1H),5.25–5.12(m ,1H),4.50(d,2H),3.96-3.67(m,3H),3.16(dd,2H),2.96-2.71(m,2H),1.76(d,2H),1.49-1.51(m,1H),0.98(s,6H).

[0117] Example 6

[0118] The synthetic route for compound C10-446 is shown below:

[0119]

[0120] 1. Synthesize compound 2A

[0121] 1A was dissolved in DCM and TFA and stirred at room temperature for 2 hours. The reaction was concentrated to give 2A. LC-MS (ESI+): m / z = 266 [M+1] + .

[0122] 2. Synthesis of compound 3A

[0123] Dissolve 2A and 2B in DMF, add DIPEA, cool to 0℃, and add dropwise 50% T4P / DMF solution (20.36 g, 28.3 mmol). After the addition is complete, stir at room temperature for 2 h. Quench with water (500 mL), extract three times with DCM (300 mL), combine the organic phases, wash with 5% citric acid aqueous solution (500 mL) and 5% sodium chloride aqueous solution (500 mL), dry to anhydrous sodium sulfate, concentrate at 35℃, and purify by column chromatography to obtain 3A. LC-MS (ESI+): m / z = 479 [M+1] + .

[0124] 3. Synthesize compound 4A

[0125] 3A was dissolved in MeOH and 36.5% hydrochloric acid with stirring. 10% palladium on carbon was added, and the mixture was purged three times with hydrogen. The reaction mixture was then reacted at room temperature for 3 hours. The reaction solution was concentrated at 35°C to obtain 4A. LC-MS (ESI+): m / z = 345 [M+H] + .

[0126] 4. Synthesis of compound 5A

[0127] 4A and 4B were dissolved in methanol, and sodium cyanoborohydride was added. The reaction mixture was reacted at room temperature for 2 hours. The reaction solution was quenched with 10% sodium bicarbonate aqueous solution, extracted twice with DCM, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI+): m / z = 610 [M+1] + .

[0128] 5. Synthesis of compound 5B

[0129] 5C, DPPA, and triethylamine were dissolved in ultra-dry toluene and reacted at 110°C for 16 hours. The reaction solution was then concentrated to obtain crude liquid 5B.

[0130] 6. Synthesize compound 6A

[0131] 5A was dissolved in THF and added dropwise to a toluene solution of 5B. The reaction was allowed to proceed at room temperature for 16 hours. The reaction was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% citric acid aqueous solution and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated at 35°C to obtain 6A. LC-MS (ESI+): m / z = 771 [M+1] + .

[0132] 7. Synthesize compound 7A

[0133] 6A was dissolved in ultradry THF (60 mL) and DCM (60 mL), and Burgess reagent was added. The reaction was allowed to proceed at room temperature for 0.5 h. The reaction was quenched with water (200 mL), extracted three times with DCM (100 mL), and the organic phases were combined, washed with 5% sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, concentrated at 35 °C, and subjected to column chromatography to obtain 7A. LC-MS (ESI+): m / z = 753 [M+1] + .

[0134] 8. Synthesize compound 8A

[0135] 7A was dissolved in DCM (15 mL) and piperidine (8 mL) and reacted at room temperature for 0.5 h. Concentration and column chromatography at 35 °C yielded 8A. LC-MS (ESI+): m / z = 531 [M+1] + .

[0136] 9. Synthesized product 9A (C10-446)

[0137] 8B and HOBT were dissolved in DMF and reacted at room temperature for 10 minutes. Then, 8A and EDCI were added and reacted at room temperature for 2 hours. The reaction was quenched with water (100 mL), extracted twice with DCM (100 mL), and the organic phases were combined. The mixture was washed with 5% citric acid aqueous solution (100 mL) and 5% sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, and column chromatography was performed to obtain C10-446. LC-MS (ESI+): m / z = 616 [M+1] + . 1 H-NMR (400MHz, DMSO-d6)δ=10.73(s,1H),8.83(s,1H),7.23(t,1H),6.95(d,1H),6.91(dd,2H),6.22(d,1H),5.11(t,1H),5.04(t,1H),4.00(d,1H) ),3.94(s,2H),3.81(d,1H),3.25-3.15(m,4H),2.83(s,6H),2.62(dd,1H ),2.02-1.64(m,6H),1.60-1.34(m,6H),1.26-1.13(m,1H),0.89(dd,6H).

[0138] Example 7

[0139] The synthetic route for compound C10-457 is shown below:

[0140]

[0141] 1. Synthetic intermediate 2A

[0142] At room temperature, 1A was dissolved in ethyl acetate, and N-hydroxysuccinimide and dicyclohexylcarbodiimide were added separately. The mixture was stirred under nitrogen for 14 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude intermediate. The crude intermediate was dissolved in dichloromethane, trifluoroacetic acid was added, and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product 2A. LCMS (ESI+): m / z = 467.2 [M+H] + .

[0143] 2. Synthetic intermediate 2B

[0144] At room temperature, 1B was dissolved in dichloromethane, piperidine was added, and the mixture was stirred for 2 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was concentrated, and the residue was purified by column chromatography to obtain 2B. Data: LCMS (ESI+): m / z = 352.4 [M+H] + .

[0145] 3. Synthetic intermediate 3A

[0146] At room temperature, 2B was dissolved in dichloromethane, N,N-diisopropylethylamine was added, and the mixture was stirred for 2 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction mixture was poured into water (10 mL), extracted with dichloromethane (15 mL * 2), and separated. The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 3A. Data: LCMS (ESI+): m / z = 882.4 [M + H] + .

[0147] 4. Synthetic product 4A (C10-457)

[0148] 3A was dissolved in dichloromethane at room temperature, piperidine was added, and the mixture was stirred at 25°C for 2 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was concentrated, and the crude product was purified by high-performance liquid chromatography and then lyophilized to obtain product C10-457. LC-MS (ESI+): m / z = 660.4 [M+H] + . 1 H NMR (400MHz, DMSO-d) 6 )δ10.73(s,1H),8.55(s,1H),7.61(brs,2H),7.27–7.20(m,1H),6.97(d,J=7.1Hz,1H),6.94–6.86(m ,2H),6.79(d,J=7.1Hz,1H),5.10(dd,J=16.0,8.3Hz,2H),3.98(d,J=10.6Hz,1H),3.76(d,J=10.6Hz ,1H),3.20(t,J=6.4Hz,1H),3.13–3.07(m,2H),3.03–2.98(m,1H),2.67–2.60(m,1H),2.48–2.43(m, 1H),2.34–2.26(m,2H),2.05–1.51(m,12H),1.40–1.28(m,2H),1.18–1.09(m,1H),0.90–0.83(m,6H).

[0149] Example 8

[0150] The synthetic route for compound C10-458 is shown below:

[0151]

[0152] The specific synthesis process is similar to that of compound C10-457. LCMS (ESI+): m / z = 660.4 [M+H]+ . 1 H NMR (400MHz, DMSO-d) 6 )δ10.70(s,1H),8.38(s,1H),7.22(dt,J=0.8Hz,J=7.6Hz,1H),6.95(d,J=7.2Hz,1H),6.89(t,J=8Hz,1 H),,6.69(d,J=7.0Hz,1H),5.15–5.05(m,2H),3.99(d,J=10.8Hz,1H),3.76(d,J=10.7Hz,1H),3.43–3. 38(m,1H),3.28–3.23(m,1H),3.17–3.04(m,4H),2.67–2.58(m,1H),2.48–2.43(m,1H),2.31(t,J=7.5H z,2H),2.04–1.96(m,1H),1.84–1.48(m,9H),1.39–1.28(m,2H),1.18–1.09(m,1H),0.91–0.84(m,6H).

[0153] Example 9

[0154] The synthetic route for compound C10-459 is shown below:

[0155]

[0156] The specific synthesis process is similar to that of compound C10-457. LCMS (ESI+): m / z = 659.4 [M+H] + . 1 H NMR (400MHz, CD3OD) δ7.29–7.23(m,1H),7.01–6.92(m,3H),5.28–5.19(m,1H),5.13(t,J=8.2Hz,

[0157] 1H),4.25(d,J=10.7Hz,1H),3.90(d,J=10.6Hz,1H),3.51–3.34(m,3H),3.27–3.21(m,1H),3.16–3.11(m,2H),2.68–2.64(m,1H),2. 38–2.35(m,1H),2.09–1.98(m,2H),1.96–1.86(m,2H),1.81–1.58(m,8H),1.53–1.42(m,2H),1.38–1.30(m,1H),1.03–0.87(m,6H).

[0158] Example 10

[0159] The synthetic route for compound C10-466 is shown below:

[0160]

[0161] 1. Synthetic intermediate 2A

[0162] At room temperature, 1A was dissolved in dichloromethane, and 1-hydroxybenzotriazole, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N-diisopropylethylamine, and aminoacetaldehyde dimethyl acetal were added, followed by stirring under nitrogen atmosphere for 6 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was poured into water, extracted with dichloromethane, and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 2A. LC-MS (ESI+): m / z = 449.2 [M+Na] + .

[0163] 2. Synthetic intermediate 3A

[0164] At room temperature, 2A was dissolved in tetrahydrofuran, and 6N hydrochloric acid was added. The mixture was stirred for 2 hours under these conditions. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was poured into water, extracted with ethyl acetate, and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude 3A. LC-MS (ESI+): m / z = 381.2 [M+H] + .

[0165] 3. Synthetic intermediate 4A

[0166] At room temperature, 3A, L-leucine tert-butyl hydrochloride, was dissolved in methanol, and sodium cyanoborohydride was added. The mixture was stirred for 16 hours under these conditions. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was poured into water, extracted with ethyl acetate, and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 4A. LCMS (ESI+): m / z = 552.4 [M+H] + .

[0167] 4. Synthetic intermediate 5A

[0168] At room temperature, 4A, 4,6,7-trifluoro-1H-indole-2-carboxylic acid, and N,N-diisopropylethylamine were dissolved in N,N-dimethylformamide, and 1-butylphosphine anhydride was added. The reaction was stirred at 40°C for 5 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was poured into water, extracted with ethyl acetate, and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI+): m / z = 771.3 [M+Na] + .

[0169] 5. Synthetic intermediate 6A

[0170] At room temperature, 5A was dissolved in dichloromethane, and trifluoroacetic acid was added. The reaction was stirred at 25°C for 4 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was monitored. The reaction solution was concentrated and freeze-dried to obtain 6A. LC-MS (ESI+): m / z = 693.1 [M+H] + .

[0171] 6. Synthetic intermediate 7A

[0172] At room temperature, 6A was dissolved in N,N-dimethylformamide, and 1-hydroxybenzotriazole was added at 0°C. The reaction mixture was stirred at 0°C for 10 minutes. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and reactant 6B were added separately, and the reaction system was stirred at 0°C for 3 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction mixture was poured into water, extracted with ethyl acetate, and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 7A. LC-MS (ESI+): m / z = 928.3 [M+Na] + .

[0173] 7. Synthetic intermediate 8A

[0174] At room temperature, 7A was dissolved in tetrahydrofuran, and Burgess reagent was added. The reaction was stirred at 25°C for 2 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was poured into water, extracted with ethyl acetate, and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 8A. LC-MS (ESI+): m / z = 888.1 [M+H] + .

[0175] 8. Synthesized product 9A (C10-466)

[0176] 8A was dissolved in dichloromethane at room temperature, piperidine was added, and the mixture was stirred at 25°C for 2 hours. LC-MS showed that the starting material was completely consumed, and the MS of the product was detected. The reaction solution was concentrated, and the residue was purified by column chromatography to obtain the crude product. The crude product was purified by high-performance liquid chromatography and lyophilized to obtain C10-466. LC-MS (ESI+): m / z = 666.3 [M+H] + . 1 HNMR (400MHz, CD3OD) δ7.21–7.07(m,2H),7.03–6.84(m,3H),6.83–6.75(m,1H),5.38(s,1H),5.20(t,J=7.9Hz,1H),4.19–3.90(m,3H) ,3.83–3.75(m,1H),3.55–3.43(m,2H),3.19(s,1H),2.68(d,J=7.9Hz,2H),2.05–1.87(m,3H),1.71–1.63(m,1H),1.12–0.84(m,12H).

[0177] Example 11

[0178] The synthetic route for compound C10-467 is shown below:

[0179]

[0180] 1. Synthetic intermediate 2A

[0181] 1A, 1B, potassium carbonate, and potassium iodide were added to acetonitrile and reacted at 80°C for 16 hours. The reaction solution was filtered and concentrated to obtain 2A. LC-MS (ESI+): m / z = 246 [M+H] + .

[0182] 2. Synthetic intermediate 3A

[0183] 2A, 2B, and DIEA were added to DMF and stirred until dissolved. HATU was added at room temperature, and the mixture was stirred at 50°C for 2 hours. The reaction solution was quenched with water, extracted three times with DCM, and the organic phases were combined. The mixture was washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 3A. LC-MS (ESI+): m / z

[0184] =443[M+H] + .

[0185] 3. Synthetic intermediate 4A

[0186] 3A was dissolved in DCM, and TFA was added at 0°C. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 hours. The reaction solution was concentrated at 35°C to obtain 4A. LC-MS (ESI+): m / z = 387 [M+H] + .

[0187] 4. Synthetic intermediate 5A

[0188] 4A was dissolved in DMF, and HOBT was added at 0°C. After the addition was complete, the mixture was stirred for 10 minutes, and then 4B and EDCI were added at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 hours. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI-): m / z = 598 [MH] - .

[0189] 5. Synthesized product 6A (C10-467)

[0190] 5A was dissolved in ultradry dichloromethane (2 mL) and ultradry tetrahydrofuran (2 mL), and Burgess reagent was added at room temperature. The reaction solution was quenched with water and dichloromethane, the layers were separated, dried, and purified by column chromatography to obtain C10-467. LC-MS (ESI-): m / z = 580 [MH] - . 1 H-NMR (400MHz, DMSO-d6) δ = 12.57 (s, 1H), 10.65 (s, 1H), 7.21-7.05 (m, 3H), 6.86 (dd, 2H), 6.64 (s, 1H), 5.22-5.15 (m, 1H), 3 .94-3.79(m,2H),3.75-3.61(m,2H),3.39(s,2H),3.18(s,3H),2.66(dd,1H),1.93-1.48(m,4H),1.25(d,1H),0.94(d,6H).

[0191] Example 12

[0192] The synthetic route for compound C10-485 is shown below:

[0193]

[0194] 1. Synthetic intermediate 2A

[0195] At 25°C, 1B and sodium cyanoborohydride were added to a MeOH solution of 1A. LCMS showed that the starting material was completely consumed, and the MS of the product was monitored. The reaction solution was purified with H₂O and extracted with DCM. The organic layers were combined, dried with anhydrous Na₂SO₄, filtered, and concentrated to obtain an oily substance 2A, which was used directly in the next reaction. LCMS (ESI+): m / z = 242.3 [M+H] + .

[0196] 2. Synthetic intermediate 3A

[0197] At 25°C, 2A and 2B were dissolved in DMF, and n-butylphosphine anhydride (50% ethyl acetate solution, 1164 mg, 3.23 mmol) and N,N-diisopropylethylamine were added. The mixture was stirred at 40°C for 2 hours. LC-MS showed that the starting materials were completely consumed, and the MS of the product was detected. The reaction was extinguished by water extraction, followed by extraction with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and obtained by column chromatography to yield 3A. LC-MS (ESI+): m / z = 460.7 [M+Na] + .

[0198] 3. Synthetic intermediate 4A

[0199] At 25°C, starting material 3A was dissolved in DCM, TFA was added, and the mixture was stirred at 25°C for 2 hours. LCMS showed that the starting material was completely consumed, and the MS of the product was monitored. The reaction solution was concentrated under reduced pressure to obtain 4A. LCMS (ESI+): m / z = 382.8 [M+H] + .

[0200] 4. Synthetic intermediate 5A

[0201] At 0°C, starting material 4A and INT-2 were dissolved in DCM, and EDCI was added. The system was slowly heated to 25°C and stirred for 1 hour. LCMS showed that the starting material was completely consumed, and the MS of the product was monitored. Water was added to the system to extinguish the reaction, followed by extraction with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and obtained by column chromatography to yield 5A. LCMS (ESI+): m / z = 618.3 [M+Na] + .

[0202] 5. Synthesized product 6A (C10-485)

[0203] Burgess reagent was added to a THF solution of 5A at 25°C. The reaction was stirred at 25°C for 2 hours. LCMS showed that the starting material was completely consumed and the MS of the product was detected. The reaction was quenched by adding water (50 mL), extracted with DCM (30 mL × 3), and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was obtained by column chromatography. The crude product was purified by high performance liquid chromatography and lyophilized to give 6A (C10-485). LCMS (ESI+): m / z = 600.3 [M + Na] + . 1 HNMR (400MHz, DMSO-d) 6 )δ12.65(s,1H),10.66(s,1H),7.14(ddd,3H),6.90(t,1H),6.85(d,1H),6.52(s,1H),5.27(s,1H),5.17(dd,1H),3.86(d,1H),3.60(d d,3H),3.27(dd,1H),2.68(dd,1H),1.92–1.82(m,1H),1.74(dd,1H),1.67–1.57(m,1H),0.93(dd,8H),0.52–0.38(m,2H),0.15(s,1H).

[0204] Example 13

[0205] The synthetic route for compound C10-515 is shown below:

[0206]

[0207] 1. Synthetic intermediate 2A

[0208] 1A, 1B, potassium carbonate, and potassium iodide were added to acetonitrile and reacted at 80°C for 16 hours. The reaction solution was filtered, concentrated, and subjected to column chromatography to obtain 2A. LC-MS (ESI+): m / z = 246 [M+H] + .

[0209] 2. Synthetic intermediate 3A

[0210] 2A, 2B, DIEA, and DMAP were added to DMF and stirred until dissolved. HATU was added at room temperature, and the mixture was stirred at 60°C for 3 hours. The reaction solution was quenched with water, extracted three times with DCM, and the organic phases were combined. The mixture was washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 3A. LC-MS (ESI+): m / z = 407 [M+H] + .

[0211] 3. Synthetic intermediate 4A

[0212] 3A was dissolved in DCM, and TFA was added at 0°C. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 hours. The reaction solution was concentrated at 35°C to obtain 4A. LC-MS (ESI+): m / z = 351 [M+H] + .

[0213] 4. Synthetic intermediate 5A

[0214] 4A was dissolved in DMF, and HOBT was added at 0°C. After the addition was complete, the mixture was stirred for 10 minutes, and then 4B and EDCI were added at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 hours. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI-): m / z = 562 [MH] - .

[0215] 5. Synthesized product 6A (C10-515)

[0216] 5A was dissolved in ultradry dichloromethane, and Burgess reagent was added at room temperature. The reaction solution was quenched with water and dichloromethane, the layers were separated, dried, and purified by column chromatography to obtain C10-515. LC-MS (ESI-): m / z = 544 [MH] - . 1 H-NMR (400MHz, DMSO-d6) δ = 11.86 (s, 1H), 10.65 (s, 1H), 7.23-7.09 (m, 4H), 6.94-6.66 (m, 4H), 5.28 (s, 1H) ,5.19(dd,1H),3.97-3.66(m,4H),3.22(s,3H),2.66(dd,1H),1.85-1.45(m,4H),1.25(d,2H),0.94(d,6H).

[0217] Example 14

[0218] The synthetic route for compound C10-525 is shown below:

[0219]

[0220] 1. Synthetic intermediate 2A

[0221] Dissolve 1A and 1B in ACN, add potassium carbonate and KI, and stir overnight at 80°C. LCMS showed complete conversion of the starting materials. Filter the reaction solution, concentrate the filtrate under reduced pressure to obtain the crude product, and purify by column chromatography to obtain 2A. LCMS (ESI+): m / z = 202.0 [M+1] + .

[0222] 2. Synthetic intermediate 3A

[0223] 2A was dissolved in THF, and LiOH dissolved in water was added. The mixture was stirred at room temperature for 1 hour. LCMS showed complete conversion of the starting material. The reaction solution was concentrated under reduced pressure to obtain 3A. LCMS (ESI-): m / z = 186.0 [M-1] - .

[0224] 3. Synthetic intermediate 4A

[0225] Dissolve 3B in DCM, add one drop of DMF, cool to 0°C, slowly add oxaloyl chloride, and stir at room temperature for 30 min. In a separate reaction flask, dissolve 3A in DCM, add TEA, cool to 0°C, slowly add the above reaction solution, and stir at room temperature for 30 min. LCMS showed complete conversion of the starting materials. Quench the reaction solution with water to extinguish the reaction, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate for 0.5 h, filter, and concentrate under reduced pressure to obtain 4A. LCMS (ESI+): m / z = 367.0 [M+1] + .

[0226] 4. Synthetic intermediate 5A

[0227] Dissolve 4A and 4B in DMF, add DIEA and T4P, stir at room temperature for 1 h, and LCMS shows complete conversion of the starting materials. Quench the reaction solution with water, extract with ethyl acetate, combine the organic phases, wash with dilute hydrochloric acid solution, wash with saturated sodium bicarbonate solution, wash with saturated brine, dry with anhydrous sodium sulfate for 0.5 h, filter, concentrate under reduced pressure to obtain crude product, purify by column chromatography, and purify by reverse column chromatography to obtain 5A. LCMS (ESI-): m / z = 578.3 [M-1] - .

[0228] 5. Synthesized product 6A (C10-525)

[0229] 5A was dissolved in DCM, Burgess reagent was added, and the mixture was stirred at room temperature for 1 hour. LC-MS showed complete conversion of the starting material. The reaction solution was purified by column chromatography to obtain 6A. LC-MS (ESI-): m / z = 560.2 [M-1] - . 1H NMR(400MHz,DMSO-d6)δ12.01(s,1H),10.65(s,1H),7.22(t,1H),7.09(d,1H),7.01–6.94(m,3H),6.88–6.83(m,2H),5.22(dd,1H),3.96–3.6 3(m,5H),3.56–3.42(m,2H),3.20(s,3H),2.61(dd,1H),2.00(dt,1H), 1.28–1.11(m,2H),0.70–0.59(m,1H),0.31(m,2H),0.11–0.02(m,2H).

[0230] Example 15

[0231] The synthetic route for compound C10-526 is shown below:

[0232]

[0233] LCMS(ESI-): m / z = 520.0 [M-1] - . 1 H NMR(400MHz,DMSO-d6)δ11.97(s,1H),10.75(s,

[0234] 1H),7.09(t,2H),7.00(d,1H),6.92–6.77(m,3H),6.73(s,1H),5.24–4.98(m,2H),3.94–3.8 2(m,2H),3.57–3.46(m,2H),3.22(s,3H),2.67–2.55(m,1H),2.52–2.42(m,3H),1.42(d,3H).

[0235] Example 16

[0236] The synthetic route for compound C10-563 is shown below:

[0237]

[0238] 1. Synthetic intermediate 2A

[0239] 1A, 1B, potassium carbonate, and potassium iodide were added to acetonitrile and reacted at 80°C for 16 hours. The reaction solution was filtered, concentrated, and subjected to column chromatography to obtain 2A. LC-MS (ESI+): m / z = 256 [M+H] + .

[0240] 2. Synthetic intermediate 3A

[0241] 2A, 2B, and DIEA were added to DMF and stirred until dissolved. A 50% T4P / DMF solution was added at room temperature, and the mixture was stirred for 3 hours at room temperature. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution. The mixture was dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 3A. LC-MS (ESI+): m / z = 379 [M-55] + .

[0242] 3. Synthetic intermediate 4A

[0243] 3A was dissolved in DCM, and TFA was added at 0°C. After the addition was complete, the reaction mixture was allowed to react at room temperature for 2 hours. The reaction solution was concentrated at 35°C to give 4A. LC-MS (ESI+): m / z = 379 [M+H] + .

[0244] 4. Synthetic intermediate 5A

[0245] 4A was dissolved in DMF, and HOBT was added at 0°C. After the addition was complete, the mixture was stirred for 10 minutes, and then 4B and EDCI were added at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 hours. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI-): m / z = 590 [MH] - .

[0246] 5. Synthesized product 6A (C10-563)

[0247] 5A was dissolved in ultra-dry dichloromethane, and Burgess reagent was added at room temperature. The reaction mixture was reacted for half an hour at room temperature. The reaction solution was quenched with water and dichloromethane, separated into layers, dried, concentrated, and purified by plate filtration to obtain C10-563. LC-MS (ESI-): m / z = 572 [MH] - .

[0248] 1 H-NMR (400MHz, DMSO-d6) δ = 12.09 (s, 1H), 10.69 (s, 1H), 7.42-6.85 (m, 7H), 5.43-5.19 (m, 2H), 3.84 -3.49(m,4H),2.80-2.58(m,2H),2.04-1.79(m,2H),1.53(ddt,4H),1.23(s,6H),0.92-0.83(m,4H).

[0249] Example 17

[0250] The synthetic route for compound C10-565 is shown below:

[0251]

[0252] 1. Synthetic intermediate 2A

[0253] Dissolve 1A and 1B in ACN, add potassium carbonate and KI, and stir overnight at 80°C. LCMS showed complete conversion of the starting materials. Filter the reaction solution, concentrate the filtrate under reduced pressure to obtain the crude product, and purify by column chromatography to obtain 2A. LCMS (ESI+): m / z = 212.0 [M+1] + .

[0254] 2. Synthetic intermediate 3A

[0255] 2A was dissolved in THF, and LiOH dissolved in water was added. The mixture was stirred at room temperature for 1 hour. LC-MS showed complete conversion of the starting material. The reaction solution was concentrated under reduced pressure to obtain 3A. LC-MS (ESI-): m / z = 196.0 [M-1] - .

[0256] 3. Synthetic intermediate 4A

[0257] Dissolve 3B in DCM, add one drop of DMF, cool to 0°C, slowly add oxaloyl chloride, and stir at room temperature for 30 min. In a separate reaction flask, dissolve 3A in DCM, add TEA, and LCMS shows complete conversion of the starting material. Quench the reaction mixture with water, extract with ethyl acetate, combine the organic phases, wash with dilute hydrochloric acid solution, wash with saturated sodium bicarbonate solution, wash with saturated brine, dry with anhydrous sodium sulfate for 0.5 h, filter, concentrate under reduced pressure to obtain crude product, and purify by column chromatography to obtain 5A. LCMS (ESI+): m / z = 367.0 [M+1] + .

[0258] 4. Synthetic intermediate 5A

[0259] 4A and 4B were dissolved in DMF, and DIEA and T4P were added. The mixture was stirred at room temperature for 1 hour, and LCMS showed complete conversion of the starting materials. The reaction solution was quenched with water, extracted with ethyl acetate, and the combined organic phases were washed with dilute hydrochloric acid solution, saturated sodium bicarbonate solution, and saturated brine solution. After drying with anhydrous sodium sulfate for 0.5 hours, the mixture was filtered, concentrated under reduced pressure to obtain the crude product, and purified by column chromatography to obtain 5A. LCMS (ESI-): m / z = 588.0 [M-1] - .

[0260] 5. Synthesized product 6A (C10-565)

[0261] 5A was dissolved in DCM, Burgess reagent was added, and the mixture was stirred at room temperature for 1 h. LC-MS showed complete conversion of the starting material. The reaction solution was purified by column chromatography to obtain C10-565. LC-MS (ESI-): m / z = 570.2 [M-1]- . 1 H NMR (400MHz, DMSO-d6) δ11.97(d,1H),10.63(s,1H),7.15(m,2H),6.95(s,2H),6.84(m,2H),6.69–6.66(m,1H),3.83(d,2H),3.67( s,3H),3.47(s,1H),2.69–2.55(m,1H),2.45(d,1H),1.98(m,1H),1.74–1.37(m,4H),1.21(d,1H),0.47–0.23(m,5H),0.10(m,3H).

[0262] Example 18

[0263] The synthetic route for compound C10-571 is shown below:

[0264]

[0265] 1. Synthetic intermediate 2A

[0266] 1A was dissolved in methanol, and thionyl chloride was added dropwise at 0°C. After the addition was complete, the temperature was slowly raised to 50°C and the reaction was carried out for 4 hours. The reaction solution was concentrated to obtain 2A hydrochloride. LC-MS (ESI-): m / z = 128 [MH] - .

[0267] 2. Synthetic intermediate 3A

[0268] 2A was dissolved in DCM, triethylamine was added, and trifluoromethanesulfonic anhydride was added dropwise at 0°C. After the addition was complete, the reaction mixture was reacted at rest for 4 hours. The reaction solution was quenched with water, and the mixture was extracted twice with DCM. The organic compounds were combined, washed with 5% hydrochloric acid and then with 5% brine, dried, and concentrated to obtain 3A. LC-MS (ESI-): m / z = 260 [MH] - .

[0269] 3. Synthetic intermediate 4A

[0270] 3A was dissolved in a mixed solution of sodium hydroxide and water, and reacted at RT for 16 h. The pH of the reaction solution was adjusted to 2 with 10% hydrochloric acid. The reaction solution was then concentrated to dryness at 50 °C. The solid was then dissolved in methanol by slurry mixing, filtered, and the filtrate was dried and concentrated to dryness. The residue after methanol concentration was dissolved in ethyl acetate, filtered to remove inorganic salts, and the ethyl acetate filtrate was concentrated to obtain 4A. LC-MS (ESI-): m / z = 246 [MH] - .

[0271] 4. Synthetic intermediate 5A

[0272] 4A, 4B, and DIEA were added to DMF and stirred until dissolved. A 50% T4P / DMF solution was added at room temperature, and the mixture was stirred for 2 hours at room temperature. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution. The mixture was dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 5A. LC-MS (ESI-): m / z = 483 [MH] - .

[0273] 5. Synthetic intermediate 6A

[0274] 5A was dissolved in DCM, and TFA was added at 0°C. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 hours. The reaction solution was concentrated at 35°C to obtain 6A. LC-MS (ESI-): m / z = 427 [MH] - .

[0275] 6. Synthetic intermediate 7A

[0276] 6A, 6B, and DIEA were added to DMF and stirred until dissolved. A 50% T4P / DMF solution was added at room temperature, and the mixture was stirred for 2 hours at room temperature. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution. The mixture was dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 7A. LC-MS (ESI-): m / z = 640 [MH] - .

[0277] 7. Synthesized product 8A (C10-571)

[0278] 7A was dissolved in ultra-dry dichloromethane, and Burgess reagent was added at room temperature. The reaction mixture was reacted for half an hour at room temperature. The reaction solution was quenched with water and dichloromethane, separated into layers, dried, concentrated, and purified by plate separation to obtain C10-571. LC-MS (ESI-): m / z = 622 [MH] - .

[0279] 1 H-NMR (400MHz, CDCl3) δ = 8.52 (s, 1H), 7.38-7.27 (m, 1H), 7.09-6.80 (m, 3H), 5.18 (t, 1H), 5.03-4.89 (m, 1H), 4.00 -3.92(m,1H),3.80-3.66(m,1H),3.49-3.37(m,1H),2.86(dd,1H),2.52(s,1H),1.25(s,19H),0.98-0.92(m,4H).

[0280] Example 19

[0281] The synthetic route for compound C10-579 is shown below:

[0282]

[0283] 1. Synthetic intermediate 2A

[0284] 1A, 1B, and DIEA were added to DMF and stirred until dissolved. A 50% T4P / DMF solution was added at room temperature, and the mixture was stirred at 50°C for 2 hours. The reaction solution was quenched with water, extracted three times with DCM, and the combined organic phases were washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution. The mixture was dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography to obtain 2A. LC-MS (ESI-): m / z = 469 [MH] - .

[0285] 2. Synthetic intermediate 3A

[0286] 2A was dissolved in DCM, and TFA was added at 0°C. After the addition was complete, the reaction mixture was allowed to react at room temperature for 2 hours. The reaction solution was concentrated at 35°C to obtain 3A. LC-MS (ESI-): m / z = 413 [MH] - .

[0287] 3. Synthetic intermediate 4A

[0288] 3A, 3B, and DIEA were added to DMF and stirred until dissolved. A 50% T4P / DMF solution was added at room temperature, and the mixture was stirred for 2 hours at room temperature. The reaction solution was quenched with water, extracted three times with DCM, and the organic phases were combined. The mixture was washed with 5% hydrochloric acid, 5% sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated at 35°C, and the residue was purified by column chromatography followed by preparative plate separation to obtain 4A. LC-MS (ESI-): m / z = 626 [MH] - .

[0289] 4. Synthesized product 5A (C10-579)

[0290] 4A was dissolved in ultra-dry dichloromethane, and Burgess reagent was added at room temperature. The reaction mixture was reacted for half an hour at room temperature. The reaction solution was quenched with water and dichloromethane, separated into layers, dried, concentrated, and purified by preparative chromatography to obtain C10-579. LC-MS (ESI-): m / z = 608 [MH] - . 1H NMR (400MHz, DMSO-d6) δ10.74(s,1H),9.85(d,1H),7.23(t,1H),7.09(d,1H),6.94(t,1H),6. 86(d,1H),5.25–5.15(m,2H),4.30-4.26(m,1H),3.84(d,1H),3.47(d,1H),3.37-3.31(m,1H) ,3.05–2.99(m,1H),2.67–2.62(m,1H),2.52–2.47(m,1H),1.84–1.77(m,1H),1.61–1.46(m,2 H),0.90(dd,6H),0.59-0.57(m,3H),0.37-0.35(m,1H),0.28-0.18(m,3H),0.09-0.07(m,1H).

[0291] Example 20

[0292] The synthetic route for compound C10-580 is shown below:

[0293]

[0294] 1. Synthetic intermediate 2A

[0295] Dissolve 1A in DCM, add one drop of DMF, cool to 0°C, slowly add oxaloyl chloride, and stir at room temperature for 30 min. In a separate reaction flask, dissolve 1B in DCM, add TEA, cool to 0°C, slowly add the above reaction solution, and stir at room temperature for 30 min. LCMS showed complete conversion of the starting material. Quench the reaction solution with water to extinguish the reaction, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate for 0.5 h, filter, and concentrate under reduced pressure to obtain 2A. LCMS (ESI+): m / z = 433.0 [M+1-BOC] + .

[0296] 2. Synthetic intermediate 3A

[0297] 2A was dissolved in DCM, TFA was added, and the mixture was stirred at room temperature for 1 hour. LCMS showed complete conversion of the starting material. The reaction solution was concentrated under reduced pressure to obtain 3A. LCMS (ESI-): m / z = 417.2 [M-1] - .

[0298] 3. Synthetic intermediate 4A

[0299] Dissolve 3A and 3B in DMF, add DIEA and T4P, stir at room temperature for 1 h, and LCMS shows complete conversion of the starting materials. Quench the reaction solution with water, extract with ethyl acetate, combine the organic phases, wash with dilute hydrochloric acid solution, wash with saturated sodium bicarbonate solution, wash with saturated brine, dry with anhydrous sodium sulfate for 0.5 h, filter, concentrate under reduced pressure to obtain crude product, purify by column chromatography, and purify by reverse column chromatography to obtain 4A. LCMS (ESI-): m / z = 630.3 [M-1] - .

[0300] 4. Synthesized product 5A (C10-580)

[0301] 5A was dissolved in DCM, Burgess reagent was added, and the mixture was stirred at room temperature for 1 hour. LC-MS showed complete conversion of the starting material. The reaction solution was purified by column chromatography to obtain C10-580. LC-MS (ESI-): m / z = 612.3 [M-1] - . 1 H NMR (400MHz, DMSO-d6) δ10.72(s,1H),9.71(s,1H),7.21(m,1H),7.04(d,1H),6.93(m,1H),6.84(m,1H),5.26–5.12(m,2H),3.82(d,1H),3.57–3. 45(m,3H),3.38(m,2H),3.22(d,4H),2.60(dd,1H),1.66(m,1H),1.50(m ,1H),1.20(d,2H),0.86(t,7H),0.19(d,2H),0.06(s,1H),-0.03(s,1H).

[0302] Example 21

[0303] The synthetic route for compound C10-587 is shown below:

[0304]

[0305] 1. Synthetic intermediate 2A

[0306] Dissolve 1A and 1B in ACN, then add K2CO3 and KI. Stir overnight at 80°C. TLC analysis showed complete conversion of the starting materials. Filter the reaction mixture and concentrate to obtain 2A. LCMS (ESI+): m / z = 198.0 [M+1] + .

[0307] 2. Synthetic intermediate 3A

[0308] Dissolve 2A and 2B in DMF, and add DIPEA and HATU at room temperature. Stir at 70°C for 2 hours. LCMS shows complete conversion of the starting materials. Quench the reaction with water, extract with DCM, combine the organic phases, wash with dilute hydrochloric acid and potassium carbonate, and concentrate to obtain 3A. LCMS (ESI+): m / z = 427.0 [M+1] + .

[0309] 3. Synthetic intermediate 4A

[0310] Dissolve 3A in MeOH and H₂O, add LiOH at room temperature, and stir for 4 hours. LCMS shows complete conversion of the starting material. Extract with EA, adjust the pH of the aqueous phase to 3-4 with dilute hydrochloric acid, then extract with DCM, and concentrate the organic phase to obtain 4A. LCMS (ESI+): m / z = 413.0 [M+1] + .

[0311] 4. Synthetic intermediate 5A

[0312] Dissolve 4A and 3B in DMF, then add DIPEA and T4P at room temperature. Stir for 1 hour at room temperature. LCMS showed complete conversion of the starting materials. Quench the reaction with water, extract with DCM, combine the organic phases, dry with anhydrous sodium sulfate, filter, concentrate, and pass through a silica gel column to obtain 5A. LCMS (ESI-): m / z = 624.3 [M-1] - .

[0313] 5. Synthesized product 6A (C10-587)

[0314] Dissolve 5A in THF and DCM, and add Burgess reagent at room temperature. Stir at room temperature for 1 hour. LCMS showed complete conversion of the starting material. Quench the reaction with water, extract with DCM, combine the organic phases, dry with anhydrous sodium sulfate, filter, concentrate, and pass through a silica gel column to give C10-587. LCMS (ESI-): m / z = 606.20 [M-1] - . 1 H NMR (400MHz, DMSO-d6) δ10.74(s,1H),9.90(d,1H),7.23(t,1H),7.08(d,1H),6. 94(t,1H),6.87(d,1H),5.19(dt,2H),4.33–4.22(m,1H),3.85(d,1H),3.61(d,1H ),3.33(dd,1H),3.15–3.04(m,2H),2.64(dd,1H),1.87(dt,1H),1.64–1.48(m,1 H),0.85(t,1H),0.69(d,2H),0.57(d,2H),0.44–0.21(m,6H),0.18–0.00(m,4H).

[0315] Example 22

[0316] The synthetic route for compound C10-588 is shown below:

[0317]

[0318] 1. Synthetic intermediate 2A

[0319] Dissolve 1A (500.00 mg, 2.02 mmol) in DCM (5 mL), add one drop of DMF, cool to 0°C, and slowly add oxaloyl chloride (256.40 mg, 2.02 mmol). Stir at room temperature for 30 min. In a separate reaction flask, dissolve 1B (398.49 mg, 2.02 mmol) in DCM (5 mL), add TEA (408.81 mg, 2.02 mmol), cool to 0°C, and slowly add the above reaction solution. Stir at room temperature for 30 min. LCMS showed complete conversion of the starting material. Quench the reaction solution with water (200 mL). Combine the organic phases and wash with saturated brine (200 mL * 2). Dry with anhydrous sodium sulfate for 0.5 h, filter, and concentrate under reduced pressure to obtain 2A. LCMS (ESI-): m / z = 425.3 [M-1] - .

[0320] 2. Synthetic intermediate 3A

[0321] 2A (700.00 mg, 1.64 mmol) and 2B (379.25 mg, 1.64 mmol) were dissolved in DMF (7 mL), and DIEA (847.81 mg, 6.56 mmol) and T4P (1774.00 mg, 2.46 mmol) were added. The mixture was stirred at room temperature for 1 h, and LCMS showed complete conversion of the starting materials. The reaction solution was quenched with water (200 mL), and extracted with ethyl acetate (100 mL * 2). The combined organic phases were washed with dilute hydrochloric acid solution (200 mL * 2), saturated sodium bicarbonate solution (200 mL * 2), and saturated brine solution (200 mL * 2). After drying with anhydrous sodium sulfate for 0.5 h, the mixture was filtered, concentrated under reduced pressure to obtain the crude product, purified by column chromatography (DCM:MeOH = 20:1), and purified by reverse-phase column chromatography to obtain 3A. LCMS (ESI-): m / z = 638.4 [M-1] - .

[0322] 3. Synthetic final product 4A (C10-588)

[0323] 3A (70.00 mg, 0.11 mmol) was dissolved in DCM (1 mL), and Burgess reagent (52.43 mg, 0.22 mmol) was added. The mixture was stirred at room temperature for 1 h, and LC-MS showed complete conversion of the starting material. The reaction solution was purified by column chromatography (PE:EA = 1:2) to obtain C10-588. LC-MS (ESI-): m / z = 620.3 [M-1] - . 1 H NMR(400MHz,DMSO-d6)δ10.73(s,1H),9.95(s,1H),7.21(td,1H),7.00(d, 1H),6.94–6.79(m,2H),5.20–5.07(m,2H),4.06(s,1H),3.88–3.69(m,2H) ,3.59–3.42(m,1H),2.59(dd,1H),2.43(d,1H),1.73(dt,1H),1.57–1.32( m,3H),1.20(s,3H),0.82(t,1H),0.47–0.29(m,4H),0.29–-0.05(m,7H),.

[0324] Example 23

[0325] The synthetic route for compound C10-595 is shown below:

[0326]

[0327] 1. Synthetic intermediate 2A

[0328] Dissolve 1A (2.00 g, 13.77 mmol) in MeOH (40 mL), cool to 0 °C, and add thionyl chloride (4.91 g, 41.31 mmol) dropwise. After the addition is complete, react at 60 °C for 4 h. Concentrate to dryness, and remove tetrahydrofuran to obtain 2A. LCMS (ESI+): m / z = 160.20 [M+H].

[0329] 2. Synthetic intermediate 3A

[0330] Add 2B (3.10g, 12.54mmol), DMF (30ml), and HATU (4.91g, 12.54mmol) sequentially to a 100mL bottle.

[0331] DIEA (8.10 g, 62.70 mmol) was added dropwise at 0℃ and stirred at room temperature for 3 hours. Then 2A (2.19 g, 13.79 mmol) was added and stirred at room temperature for 16 hours. LCMS analysis showed that the starting material reaction was complete. The reaction was stopped, and the reaction solution was quenched with water (200 mL). Extraction was performed with ethyl acetate (200 mL x 2). The organic phase was washed with 1N hydrochloric acid (200 mL), saturated sodium bicarbonate (200 mL), and saturated brine (200 mL). The organic phase was concentrated to obtain 3A. LCMS (ESI+): m / z = 389.2 [M+H]

[0332] 3. Synthetic intermediate 4A

[0333] Dissolve 3A (4.60 g, 11.84 mmol) in MeOH (20 mL), add an aqueous solution of Lithium hydroxide hydrate (1.99 g, 47.36 mmol) (20 mL) at 0 °C, and react at 25 °C for 1 h. LCMS showed that the starting material reacted completely and the product was formed. Stop the reaction, concentrate off the methanol, and extract the remaining aqueous solution with EA (50 mL). Adjust the pH of the aqueous phase to 4 with 1 N HCl, then extract with EA (50 mL). Dry the organic phase with anhydrous sodium sulfate, filter, and concentrate to obtain 4A. LCMS (ESI+): m / z = 375.2 [M+H].

[0334] 4. Synthetic intermediate 5A

[0335] Dissolve 4A (4.40 g, 11.75 mmol) and 4B (4.08 g, 17.63 mmol) in DMF (40 mL), add DIEA (7.59 g, 58.75 mmol), then add 50% T4P / DMF solution (12.69 g, 17.63 mmol) in an ice bath at 0 °C, and react at 25 °C for 1 hour. LCMS showed that the starting material reacted completely. Stop the reaction. Quench the reaction with water (400 mL), extract with EA (400 mL * 3), combine the organic phases and wash with dilute hydrochloric acid (1 N, 250 mL * 2), saturated sodium bicarbonate (250 mL * 2), and sodium chloride (250 mL). Concentrate the organic phase to obtain 6.2 g of crude product. Prepare the crude product by reverse phase (20% ACN:

[0336] The product was dissolved in an aqueous solution of H₂O / 0.1% HCOOH (80% ACN: H₂O / 0.1% HCOOH). Acetonitrile was removed by concentration, and the aqueous solution was extracted with ethyl acetate and concentrated. The resulting mixture was then separated into 50 ml of ethyl acetate and 50 ml of water. The organic phase was dried, filtered, and concentrated to obtain 5A. LCMS (ESI-): m / z = 586.3 [MH]

[0337] 5. Synthetic final product 6A (C10-595)

[0338] 5A (2.2 g, 3.74 mmol) was dissolved in DCM (24 mL) and THF (8 mL), and Burgess reagent (1.34 g, 5.61 mmol) was added. The mixture was reacted at room temperature for 8 hours. LCMS showed that 23% of the starting material remained unreacted, while the product comprised 73%. The reaction was stopped, the mixture was concentrated to dryness, and purified by column chromatography (Hex:EA = 1:1) to obtain product C10-595. LCMS (ESI-): m / z = 568.3 [MH]. 1 H NMR (400MHz, DMSO-d6) δ10.73(s,1H),9.89(d,J=8.7Hz,1H),7.23(td,J=7.7,1.3Hz,1H),7.04(dd,J=7.5,1.3Hz,1H),6.94(td ,J=7.5,1.0Hz,1H),6.90–6.84(d,1H),5.22(dd,J=9.2,5.8Hz,1H),5.14(dd,J=8.6,7.3Hz,1H),4.09(dd,J=8.7,6.8Hz,1H),3. 85(d,J=10.4Hz,1H),3.64(d,J=10.3Hz,1H),2.93(s,3H),2.63(dd,J=13.2,8.7Hz,1H),2.50–2.42(m,1H),1.60(qdd,J=14.2, 8.9,5.4Hz,2H),1.45–1.32(m,1H),0.93–0.81(m,6H),0.69(tdd,J=8.2,6.6,3.9Hz,1H),0.34–0.15(m,3H),0.15–0.07(m,1H).

[0339] Pharmacological Examples

[0340] Pharmacological Example 1: Enzyme Activity Assay: Evaluation of the in vitro anti-novel coronavirus 3CLpro protease activity of the test compound.

[0341] 1. Experimental materials:

[0342] 1.1 Reagents and Consumables and Their Sources:

[0343] Dimethyl sulfoxide (DMSO)

[0344] 96well plate

[0345] Novel Coronavirus 3CLpro Inhibitor Screening Kit

[0346] 1.2 Instruments and their sources:

[0347] Agilent Multifunctional Microplate Reader

[0348] benchtop high-speed centrifuge

[0349] 2. Experimental Methods:

[0350] The compound was dissolved in DMSO and serially diluted according to the required concentration, resulting in 10 concentration points, with each concentration in duplicate, and added to a 96-well plate. The substrate and 3CLpro protein were diluted with test buffer, and 3CLpro was added to the 96-well plate. The plate was incubated with the compound at room temperature for 10 min, followed by the addition of the substrate. The plate was then incubated at room temperature in the dark for 5-20 min, and finally, E2 was detected using a microplate reader. X / E M =340nM / 490nM. Simultaneous detection of substrate and 3CLpro protein, and background control wells containing substrate but without 3CLpro protein.

[0351] 3. Data Analysis:

[0352] (1) Calculate the inhibition rate using the following formula:

[0353] Inhibition rate % = (RFU) 100％酶活性对照 -RFU 样品 ) / (RFU 100％酶活性对照 -RFU 空白对照 )×100%;

[0354] RFU 100％酶活性对照 : Control with no inhibitory effect, containing fluorescent substrate + 3CLpro protein + 3CLpro protease, without the target compound;

[0355] RFU 样品 : Test compound wells contain fluorescent substrate + 3CLpro + 3CLpro protease + test compound;

[0356] RFU 空白对照 : 100% inhibition control, containing fluorescent substrate + 3CLpro, without 3CLpro protease, and without the test compound;

[0357] (2) The inhibition rate data (inhibition rate%) of the test compounds were analyzed using Graphpad Prism software using log(inhibitor) vs. response-variable slope (four parameters) nonlinear fitting to obtain the IC50 of the compounds. 50 The values ​​and experimental results are shown in Table 1.

[0358] Where A is less than 0.1 μm, B is 0.1-0.5 μm, C is greater than or equal to 0.5 μm and less than 1 μm, and D is greater than or equal to 1 μm.

[0359] Table 1. Results of enzyme activity testing

[0360]

[0361]

[0362] Pharmacological Example 2: ICR Mouse PK Properties Test Experiment of Compound

[0363] Referring to the PK property test experiment of ICR mice in Chinese patent application CN117986260A, the ICR mice were administered the drug at a dose of 200 mg / kg or 20 mg / kg via gavage. The solvent was 5% dimethyl sulfoxide + 5% Solutol HS15 + 5% PEG + 85% physiological saline. Blood was collected via the submandibular vein at a rate of 0.03 mL / time point. Samples were placed in tubes containing K2-EDTA and stored on ice until centrifugation. Blood samples were centrifuged at 6800 g for 6 minutes at 2–8 °C within 1 hour of collection and then frozen at approximately -80 °C. Aliquots of 10 μL plasma samples were precipitated for protein using 190 μL of 0.1% FA / CAN (containing 20 ng / mL IS). The mixture was vortexed for 3 minutes and centrifuged at 2200 rcf for 15 minutes. 120 μL of the supernatant was injected for LC-MS / MS analysis. The test results are shown in Table 2. The results show that the compound of the present invention has a high blood drug exposure in mice.

[0364] Among them, compound CN117986260A-17 in Table 2 is compound 17 in Chinese patent application CN117986260A.

[0365] Table 2. Results of ICR mouse PK property tests on the compounds.

[0366]

[0367] Pharmacological Example 3: Caco-2 Permeability Experiment

[0368] (1) Caco-2 is converted to 1×10 5 cells / cm 2The cells were seeded at a density of 10 mM in 96-transwell plates, and the culture medium was changed every 3-4 days until a confluent cell monolayer was formed in 14-28 days. The transepithelial resistance (TEER) measured by a resistance meter was higher than 400 Ω to carry out the experiment; (2) The test compound was diluted from 10 mM stock solution to 10 μM using HBSS buffer (0.5% BSA), and added to the top or outer side of the cell monolayer of the corresponding well, respectively, and incubated at 37°C and 5% BSA. (2) Incubate at a relative humidity of 95% for 120 min; (3) After incubation, collect 50 μL of the upper layer (above the cells) and the lower layer (through the cell layer into the pore membrane below) respectively, add 200 μL of ACN:MeOH (1:1) containing IS, vortex at 600 rpm for 10 min, centrifuge at 6000 rpm for 20 min, take 100 μL of supernatant from each well and transfer it to a 96-well sample plate containing 100 μL of ultrapure water for LC / MS / MS analysis, calculate the membrane permeability, efflux rate and recovery rate of the compound, and the test results are shown in Table 3.

[0369] Table 3. Results of Caco-2 Permeability Test

[0370]

[0371] Those skilled in the art will readily understand that the above description is merely an embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. Compounds of formula (I) and their pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof: in: The R1 is either absent or contains a deuterium or halogen atom, or is a C1-C4 alkyl group, or a C3-C6 cycloalkyl group, or a -C0-C4 alkylsilyl group; The R2 is selected from hydrogen, deuterium, C1-C4 alkyl, -(C0-C4 alkyl)-(C3-C6 cycloalkyl), -C0-C4 alkylsilyl, and -(C0-C4 alkyl)-phenyl; the C1-C4 alkyl, -C0-C4 alkylsilyl, and -(C0-C4 alkyl)-phenyl have one or more substituents selected from alkyl, amino, oxygen, alkoxy, hydroxyl, carboxyl, and halogen atoms; The R3 is selected from the halogenated atom, C3-C6 cycloalkyl, hydroxyl, amino, C1-C6 alkylamine, C1-C6 alkoxy, C1-C 18 Acyloxy group, C1-C 18 amide group, C1-C 18 A C1-C6 alkyl group substituted with one or more substituents in the oxycarbonyl group, wherein the substituents are optionally further replaced by an amino group, a hydroxyl group, a carboxyl group, or a (C1-C6 alkyl) group. m Amine group, (C1-C6 alkyl group) m Ammonium, C6-C 20 aryl-substituted C1-C6 alkoxy, C6-C 20 Heteroaryl-substituted C1-C6 alkoxy groups, -(OC2H4) n One or more of O (C1-C6 alkyl) are substituted; Where m is selected from 1 to 3, and n is selected from 1 to 10; The R4 is selected from NHR5, -CHR2NHSO2-(C1-C3 alkyl), -CHR2NHSO2-(C1-C3 haloalkyl), -CHR2NHC(=O)-O-(C1-C3 alkyl), OR5, or R5, wherein R5 is selected from the following groups, and R5 is optionally substituted:

2. The compound of formula (I) according to claim 1, and its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof, characterized in that, The R1 is either absent or contains deuterium or halogen atoms.

3. The compound of formula (I) according to claim 1, and its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof, characterized in that, R1 does not exist.

4. The compound of formula (I) according to claim 1, and its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof, characterized in that, R2 is selected from C1-C4 alkyl, -(C0-C4 alkyl)Si(CH3)3, -(C0-C4 alkyl)-phenyl, and -(C0-C4 alkyl)-(C3-C6 cycloalkyl).

5. The compound of formula (I) according to claim 1, and its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof, characterized in that, The R2 is selected from C1-C4 alkyl, -(C1-C2 alkyl)Si(CH3)3, and -(C1-C3 alkyl)-cyclopropyl.

6. The compound of formula (I) according to claim 1, and its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof, characterized in that, The R3 is selected from the following substituents:

7. The compound of formula (I) according to claim 1, and its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof, characterized in that, R4 is selected from 8. The following compounds and their pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers:

9. The use of the compound according to any one of claims 1-8, its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof in the preparation of medicaments for the treatment or prevention of viral infections, characterized in that, The viral infection includes viral infections selected from small RNA viruses and coronaviruses; preferably, the virus is a coronavirus selected from 229E, NL63, OC43, HKU1, SARS-CoV, MERS, or SARS-CoV-2 coronavirus.

10. The use of the compound according to any one of claims 1-8, its pharmaceutically acceptable salts, esters, prodrugs, solvates, and isomers thereof in the preparation of medicaments for treating respiratory diseases, characterized in that, The respiratory diseases include acute asthma, lung diseases secondary to environmental exposure, acute lung infections, and chronic lung infections.

11. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the compound according to any one of claims 1-8, and a pharmaceutically acceptable carrier or excipient.

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