CN116102537A - Quinolinone derivative and preparation method and application thereof - Google Patents

Abstract

The invention provides a quinolinone derivative, which has a structure shown in a formula I. It has excellent anti-Mycobacterium tuberculosis activity as a componentThe novel DprE1 enzyme inhibitor has controllable toxicity, excellent antitubercular activity and pharmacokinetic property and good clinical application prospect.

Description

A quinolinone derivative and its preparation method and use

Technical Field

The invention belongs to the field of chemical medicine, and specifically relates to a quinolinone derivative and a preparation method and use thereof.

Background Art

Tuberculosis is a chronic infectious disease caused by infection with Mycobacterium tuberculosis and is one of the top ten deadly diseases in the world. One quarter of tuberculosis deaths are caused by drug-resistant tuberculosis. So far, the treatment of drug-sensitive tuberculosis still uses four drugs from the 1970s: isoniazid, rifampicin, pyrazinamide and ethambutol. Since 2012, bedaquiline and delamanid have been approved for marketing by the US FDA and the European EMA, but due to the unclear toxicity and efficacy of bedaquiline and delamanid, they are currently controversial and are still not used as first-line anti-tuberculosis drugs. Therefore, the development of anti-Mycobacterium tuberculosis drugs with new chemical structures and modes of action is considered to be an important research direction at present.

Mycobacterium tuberculosis has a characteristic, complex cell wall structure that involves multiple functions related to cell physiology and pathogenesis. The key enzyme DprE1, which is unique to mycobacteria, is related to the biosynthesis of the cell wall of Mycobacterium tuberculosis and is essential for the survival of Mycobacterium tuberculosis. In the past few years, DprE1 enzyme inhibitors (such as BTZ and OPC-167832) have been discovered that can cause mycobacterial death by covalently or non-covalently binding to the DprE1 enzyme. These compounds have shown anti-tuberculosis efficacy, providing strong evidence for the design of DprE1 enzyme inhibitors and the treatment of tuberculosis.

However, the anti-tuberculosis activity and other properties of existing DprE1 enzyme inhibitors still need to be further improved. It is of great significance to further study quinolinone compounds with controllable toxicity, better anti-tuberculosis activity, better pharmacokinetic properties and better drug development prospects in order to obtain better clinical treatment effects.

Summary of the invention

The object of the present invention is to provide a novel DprE1 enzyme inhibitor having excellent anti-tuberculosis activity.

The present invention provides a quinolinone derivative represented by formula I or a pharmaceutically acceptable salt or solvate thereof:

Wherein, Ring A is an aryl group, an aromatic heterocyclic group or a paraaromatic heterocyclic group; L is O or NR ₇ ; L' and L" are independently selected from none, CO, SO ₂ , NH or (CH ₂ ) _m , m is an integer of 1 to 3; M ₁ and M ₂ are independently selected from N or CH;

R ₁ , R ₂ , and R ₃ are independently selected from H, halogen, C1-C3 alkoxy, hydroxyl, -NR ₁₁ R ₁₂ , or R ₂ and R ₃ are connected to form a 3-7 membered ring; R ₁₁ is H or C1-C3 alkyl, R ₁₂ is C1-C3 alkyl or -CO-R ₁₃ , and R ₁₃ is halogen substituted or unsubstituted C1-C3 alkyl;

_R4 is -H or -OH;

t为1～3的整数；R ₅ and R ₆ are independently selected from -H, -Boc _, a bridged hydrocarbon group, Ra, _Rb , _Rc substituted or unsubstituted aryl, halogen substituted or unsubstituted aromatic heterocyclic group, _Rd substituted or unsubstituted C1-C4 alkyl, C3-C6 cycloalkyl or

t is an integer from 1 to 3;

其中，R’、R”分别独立选自：H、-CO-R_s或-SO₂-R_s；R”’为-O-R_s或

R_s、R_p、R_q分别独立选自H、C1～C3的烷基，或R_p、R_q连接成环；wherein _Ra , _Rb , and _Rc are independently selected from halogen, nitro,

Wherein, R', R" are independently selected from: H, -CO- _Rs or _{-SO2- Rs} ; R"' is _-ORs or

R _s , R _p , and R _q are independently selected from H, C1-C3 alkyl, or R _p and R _q are connected to form a ring;

R _d is halogen, halogen-substituted or unsubstituted C1-C3 alkyl, C3-C6 cycloalkyl or C3-C6 heterocycloalkyl;

_R7 is a C1-C3 alkyl group.

Furthermore, the above ring A is

Furthermore, the above R ₁ , R ₂ , and R ₃ are independently selected from H, F, methoxy, hydroxyl, -NR ₁₁ R ₁₂ , or R ₁ and R ₂ are connected to form a three-membered ring; R ₁₁ is H or methyl, R ₁₂ is methyl or -CO-R ₁₃ , and R ₁₃ is methyl substituted or unsubstituted with 1 to 3 F.

Furthermore, R ₁ is H, F or methoxy, R ₂ is H or -NR ₁₁ R ₁₂ , R ₃ is H, or R ₂ and R ₃ are connected to form a saturated 3-membered carbon ring; R ₁₁ is H or methyl, R ₁₂ is methyl or -CO-R ₁₃ , and R ₁₃ is methyl or -CF ₃ .

Furthermore, the above-mentioned quinolinone derivatives have a structure shown in Formula II-A:

Preferably, R ₁ is H or F, R ₂ is H and R ₃ is H, or R ₂ and R ₃ are connected to form a saturated 3-membered carbon ring.

Furthermore, the above-mentioned quinolinone derivatives have a structure shown in Formula II-B:

wherein R ₃ is H;

Preferably, R ₁ is H or F, R ₂ is -NR ₁₁ R ₁₂ ; R ₁₁ is H or methyl, R ₁₂ is methyl or -CO-R ₁₃ , and R ₁₃ is methyl or -CF ₃ .

Furthermore, the above-mentioned quinolinone derivatives have a structure shown in Formula II-C or Formula II-D:

Wherein, R ₂ and R ₃ are both H;

Preferably, R ₁ is H, F or methoxy.

Furthermore, the above-mentioned quinolinone derivatives have a structure shown in Formula II-E or Formula II-F:

Wherein, R ₁ , R ₂ , and R ₃ are all H.

Furthermore, the above L is O or N—CH ₃ .

Furthermore, the above L' is CO, SO ₂ , NH or CH ₂ .

Furthermore, the above-mentioned L" is none.

Furthermore, at least one of the above-mentioned _M1 and _M2 is CH.

Furthermore, the above-mentioned _M1 is CH, and _M2 is N.

Furthermore, the above-mentioned _M1 is CH, and _M2 is CH.

Furthermore, the above-mentioned _M1 is N, and _M2 is CH.

Furthermore, the above R ₅ is -H or an aryl group substituted by _Ra , _Rb or _Rc .

Furthermore, the above-mentioned _Ra , _Rb , and _Rc are halogen.

Furthermore, the above _Ra , _Rb , and _Rc are independently selected from F or Cl; preferably, _Ra and _Rb are F, and _Rc is Cl.

Furthermore, the above R ₅ is -H or

t为1～2的整数。Furthermore, the above _R6 is -H, -Boc, a polycyclic bridged hydrocarbon group, an aryl group substituted by Ra, _{Rb ,} or _Rc , a nitrogen aromatic heterocyclic group substituted by two Fs, a C1-C4 alkyl group substituted or unsubstituted by _Rd , a C5-C6 cycloalkyl group, or

t is an integer from 1 to 2.

其中，R’、R”分别独立选自：H、-CO-R_s或-SO₂-R_s；R”’为-O-R_s或

R_s、R_p、R_q分别独立选自H、甲基，或R_p、R_q连接成三元环；Furthermore, the above _Ra and _Rb are F, and _Rc is selected from Cl, nitro,

Wherein, R', R" are independently selected from: H, -CO- _Rs or _{-SO2- Rs} ; R"' is _-ORs or

R _s , R _p , and R _q are independently selected from H, methyl, or R _p , R _q are connected to form a three-membered ring;

The R _d is a Cl, F substituted or unsubstituted C1-C3 alkyl, C3-C6 cycloalkyl or C6 heterocycloalkyl, wherein the heteroatom of the heterocycloalkyl is N and/or O.

Furthermore, the above R ₆ is: -H, -Boc,

Furthermore, the above-mentioned quinolinone derivatives have a structure shown in Formula III-A or Formula III-B:

Furthermore, the above-mentioned quinolinone derivatives have the following structure:

Furthermore, the above-mentioned quinolinone derivative has a structure shown in Formula III-C:

Furthermore, the above-mentioned quinolinone derivatives have the following structure:

Furthermore, the above-mentioned quinolinone derivative has a structure shown in Formula III-D:

Furthermore, the above-mentioned quinolinone derivatives have the following structure:

The present invention also provides a method for synthesizing the above-mentioned quinolinone derivatives, comprising the following steps: reacting compound A and compound B under the action of an inorganic base at 70-90° C. for 4-8 hours to obtain a compound of formula I;

The reaction formula is as follows:

Wherein, R ₀ is -OH or -NH-R ₇ .

The present invention also provides the use of the above-mentioned quinolinone derivatives or pharmaceutically acceptable salts or solvates thereof as DprE1 enzyme inhibitors.

The present invention also provides the use of the above-mentioned quinolinone derivatives or pharmaceutically acceptable salts or solvates thereof in the preparation of drugs for treating tuberculosis. Preferably, the tuberculosis is drug-resistant tuberculosis.

Furthermore, the above-mentioned drug is an anti-Mycobacterium tuberculosis drug, and preferably, the Mycobacterium tuberculosis is a drug-resistant Mycobacterium tuberculosis.

Beneficial effects of the present invention: The compound of the present invention has excellent anti-tuberculosis Mycobacterium tuberculosis activity. As a new type of DprE1 enzyme inhibitor, the toxicity is controllable, the anti-tuberculosis activity and pharmacokinetic properties are excellent, and it has very good clinical application prospects.

In the present invention, "substitution" means that one, two or more hydrogen atoms in a molecule are replaced by other different atoms or molecules, including one, two or more substitutions on isotopic atoms or heterotopic atoms in the molecule.

In the present invention, the minimum and maximum values of the carbon atom content in the hydrocarbon group are indicated by prefixes, for example, the C1-C6 alkyl or C1-6 alkyl refers to C1, C2, C3, C4, C5, C6 alkyl, i.e., a straight or branched alkyl having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, hexyl, etc. Similarly, C1-C6 alkoxy refers to C1, C2, C3, C4, C5, C6 alkoxy.

In the present invention, "salt" refers to the acidic and/or basic salts formed by a compound or its stereoisomer with an inorganic and/or organic acid and/or base, including zwitterionic salts (inner salts), and also quaternary ammonium salts, such as alkylammonium salts. These salts can be directly obtained in the final separation and purification of the compound. It can also be obtained by mixing the compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after solvent evaporation, or be obtained by freeze drying after reaction in an aqueous medium. The salt described in the present invention can be a metal ion salt (sodium salt, potassium salt, etc.) hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

In the present invention, "solvate thereof" refers to a solvate formed by a compound and a solvent, wherein the solvent includes (but is not limited to): water, ethanol, methanol, isopropanol, propylene glycol, tetrahydrofuran, and dichloromethane.

In the present invention, "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and/or the formed salt is usually chemically or physically compatible with other ingredients constituting a pharmaceutical dosage form and physiologically compatible with the receptor.

"Halogen" is fluorine, chlorine, bromine or iodine.

"Alkyl" is a hydrocarbon group formed by removing a hydrogen atom from an alkane molecule, such as methyl-CH ₃ , ethyl-CH ₃ CH ₂ , propyl, isopropyl, and the like.

"Aryl" refers to an all-carbon monocyclic or fused polycyclic group having a conjugated π-electron system, for example, phenyl, naphthyl.

"Aromatic heterocyclic group" refers to a monocyclic or fused polycyclic ring containing one to multiple heteroatoms with a conjugated π electron system. It contains at least one ring heteroatom selected from N, O or S, and the remaining ring atoms are C, and it also has a completely conjugated π electron system. For example, benzopyrazinyl.

The "aromatic heterocyclic group" refers to a group formed by the above-mentioned aryl group or aromatic heterocyclic group and another cyclic structure sharing two adjacent carbon atoms or hetero atoms.

"Heterocycle" refers to a saturated or unsaturated cyclic hydrocarbon, which may be monocyclic or polycyclic, and carries at least one ring heteroatom (including but not limited to O, S or N), and the rest are C. For example, "3-8 membered heterocycle" refers to a heterocycle having a total of 3 to 8 carbon atoms and heteroatoms.

"Cycloalkyl" refers to a saturated cyclic hydrocarbon substituent; the cyclic hydrocarbon may be a single ring or a polycyclic ring. For example, "3-8 membered cycloalkyl" refers to a cycloalkyl group having 3 to 8 carbon atoms.

"Heterocycloalkyl" refers to a saturated cyclic hydrocarbon substituent; the cyclic hydrocarbon may be monocyclic or polycyclic and carry at least one ring heteroatom (including but not limited to O, S or N). For example, "3-8 membered heterocyclic group" refers to a heterocyclic group having a total of 3 to 8 carbon atoms and heteroatoms.

"Bridged hydrocarbons" can be divided into bicyclic bridged hydrocarbons and polycyclic bridged hydrocarbons. Bicyclic bridged hydrocarbons are composed of two alicyclic rings sharing two or more carbon atoms; polycyclic bridged hydrocarbons are composed of three or more alicyclic rings sharing two or more carbon atoms. "Bridged hydrocarbon group" is a hydrocarbon group formed by missing one hydrogen atom in the molecule of a bridged hydrocarbon; "polycyclic bridged hydrocarbon group" is a hydrocarbon group formed by missing one hydrogen atom in the molecule of a polycyclic bridged hydrocarbon.

Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention.

DETAILED DESCRIPTION

The raw materials and equipment used in the present invention are all known products, which are obtained by purchasing commercially available products.

Embodiment 1,

Compound A1 5-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

The synthetic route of compound A1 is as follows:

first step:

Add 250mL of dry tetrahydrofuran to a three-necked flask, dissolve raw material 1 (28.23g, 200mmol) and raw material 2 (35.56g, 240mmol) in it, and cool the mixed system to 0℃ with an ice-salt bath. Slowly drop 110mL of 2mol/L sodium hexamethyldisilazide in tetrahydrofuran solution in a constant pressure dropping funnel. The reaction is exothermic, and the drop rate is controlled to keep the reaction temperature at 0-2℃. After the drop is completed, the reaction mixture is kept warm at 0℃ and stirred for 10min, then the ice-salt bath is removed and allowed to naturally warm to room temperature and stirred at room temperature for 3h. TLC monitors the completion of the reaction, and the reaction solution is vortexed to remove most of the solvent under reduced pressure. Ice water is added to the residue, and saturated citric acid aqueous solution is slowly added to carefully adjust the system to neutrality, then dichloromethane is added for extraction three times, the organic phases are combined, and washed with water and saturated brine in turn. The organic phase was separated and dried over anhydrous sodium sulfate, filtered and the filtrate was dried by spin drying, and purified by column chromatography to obtain 43.2 g (yield 84%) of the target intermediate 3 as a light yellow oil.

Step 2:

Add 135mL of concentrated sulfuric acid to a two-necked flask and cool the sulfuric acid to 0°C with an ice-salt bath. Dissolve intermediate 3 (43.0g, 167mmol) in 40mL of dichloromethane and slowly drip the resulting solution into the above sulfuric acid solution through a dropping funnel over 30min. Control the dripping speed to keep the temperature of the reaction liquid at 0-5°C. After the dripping is completed, the reaction mixture is kept warm at 0°C and stirred for 10min, then the ice-salt bath is removed and the mixture is allowed to naturally warm to room temperature and stirred at room temperature for 2h. TLC monitors the completion of the reaction, and the reaction solution is vortexed at 30°C under reduced pressure to remove most of the low-boiling solvent dichloromethane, and the residue is added to crushed ice. Stir the mixture until a large amount of solids precipitate, let it stand for 20min, and filter out the solids with a baffle funnel. Transfer the solids to a beaker and add water to wash them thoroughly. When the pH value of the filtrate is close to 7, the filter cake is close to light yellow to off-white. The filter cake was taken out and dried in a vacuum drying oven with phosphorus pentoxide to obtain 28.7 g (yield: 89%) of light yellow solid intermediate 4.

Step 3:

Dissolve 23.0g of intermediate 4 in 100mL of glacial acetic acid, stir and dissolve, then add 4.6g of 10% palladium carbon. Replace the nitrogen 5 times and then replace it with hydrogen once, heat to 110℃ in a hydrogen atmosphere and react for 3d. After the reaction is completed by TLC detection, the acetic acid solvent is evaporated under reduced pressure, the reaction system is diluted with PE:EA＝2:1, first filtered with diatomaceous earth, the filtrate is dried by spin-drying, and sodium bicarbonate or sodium carbonate aqueous solution is added to adjust the system pH to about 7. Ultrasonicate the mixed solution for 10min, filter the filter cake by suction, and wash the filter cake with water several times. Filter the filter cake by suction and dry it, then rinse the filter cake with DCM several times to obtain the filtrate. The filtrate is spin-dried and separated by silica gel column chromatography to obtain intermediate 521.6g, with a yield of 92.9%.

Step 4:

Weigh 21.0g of intermediate 5, dissolve it in 300mL of dry dichloromethane, cool the mixed solution to -15℃ in a -30℃ cold bath, and slowly add 400mL of BBr ₃ under this condition. Control the dropping speed to keep the system temperature below -10℃. After the addition, keep stirring at low temperature for 30min, remove the cold bath and naturally warm the reaction system to room temperature and stir for 3h. TLC monitoring shows that the reaction is very thorough. The system is spun off the solvent and low-boiling substances at room temperature under reduced pressure, and the resulting residue is added to 200mL of crushed ice and stirred until the crushed ice is completely dissolved. Add dichloromethane to extract and separate the liquids. The aqueous phase is extracted with dichloromethane three times, the organic phases are combined, and washed with water and saturated brine. Separate the organic phase and dry it with anhydrous sodium sulfate, filter and spin dry the filtrate, mix the sample column chromatography and purify to obtain 16.2g of white solid intermediate 6, with a yield of 83%.

Step 5:

Intermediate 6 (100 mg, 0.55 mmol) was dissolved in 6 mL of anhydrous DMF, anhydrous potassium carbonate (114 mg, 0.825 mmol) was added, and raw material 7 (171 mg, 0.66 mmol) was slowly added after stirring at room temperature for 5 min, and the temperature was raised to 80°C for 6 h. TLC monitored the reaction to be complete, the reaction solution was cooled to room temperature, an appropriate amount of ice water was added, and it was extracted three times with ethyl acetate, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain the target compound A1 as a light yellow solid of 24 mg, with a yield of 10%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.00(s,1H),7.34–7.19(m,2H),7.00(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.7Hz,1H),4.69(s,1H),3.77(s,2H),3.38(m,J＝10.5 ,9.1,4.6Hz,2H),3.03–2.87(m,4H),2.46(s,2H),1.79(dt,J＝12.3,6.4Hz,2H),1.63(d,J＝12.9Hz,2H).HR-MS: m/z 463.1015(M+Na) ⁺ .

Embodiment 2,

Compound A2 5-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

Referring to the synthesis method of compound A1, raw material 1 was replaced with 3-methoxyaniline to obtain compound A2, a white solid, with a yield of 15%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.01(s,1H),7.26(d,J＝8.8Hz,2H),7.07(t,J＝8.1Hz,1H),6.59(d,J＝8.2Hz,1H),6.48(d,J＝7.9Hz,1H),4.69(s,1H),3.78(s,2H ), 3.36 (d, J = 12.0Hz, 2H), 2.97 (d, J = 11.7Hz, 2H), 2.87 (t, J = 7.7Hz, 2H), 2.42 (dd, J = 8.4, 6.9Hz, 2H), 1.81 (dt, J = 12.4, 6.4Hz, 2H), 1.63 (d, J = 12.8Hz, 2H). HR-MS:m/z 445.1107(M+Na) ⁺ .

Embodiment 3,

Compound A3 4-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)quinolin-2(1H)-one

Using 4-hydroxyquinoline-2(1H)-one and raw material 7 as raw materials, referring to the fifth step synthesis method of compound A1, compound A3 was obtained as a light yellow solid with a yield of 20%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ11.33(s,1H),7.95(d,J＝8.1Hz,1H),7.51(t,J＝7.6Hz,1H),7.28(d,J＝8.7Hz,2H),7.18(t,J＝7.7Hz,1H),5.87(s,1H),4.89(s, 1H), 3.95 (s, 2H), 3.41 (d, J = 11.4Hz, 2H), 3.01 (d, J = 11.5Hz, 2H), 1.87–1.66 (m, 4H). HR-MS: m/z443.0947 (M+Na) ⁺ .

Embodiment 4,

Compound A4 4-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoroquinolin-2(1H)-one

Compound A4 was obtained by using 8-fluoro-4-hydroxyquinolin-2(1H)-one and 7 as raw materials and referring to the fifth step of the synthesis method of compound A1. The compound A4 was a pale yellow solid with a yield of 18%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.38 (s, 1H), 7.79 (d, J = 8.1 Hz, 1H), 7.44 (dd, J = 11.1, 8.0 Hz, 1H), 7.28 (d, J = 8.8 Hz, 2H), 7.18 (td, J = 8.1, 4.9 Hz, 1H), 4.92 (s, 1H), 3.97 (s, 2H), 3.00 (d, J = 11.5 Hz, 2H), 2.00 (q, J = 7.3 Hz, 2H), 1.76 (q, J = 13.3, 12.0 Hz, 4H).

Embodiment 5,

Compound A5 N-(3-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)phenyl)-2,2,2-trifluoro-N-methylacetamide

Compound A5 was obtained by using 2,2,2-trifluoro-N-(3-hydroxyphenyl)-N-methylacetamide and 7 as raw materials and referring to the fifth step of the synthesis method of compound A1. The compound A5 was a reddish brown oil with a yield of 15.3%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ7.38(t, J＝8.1 Hz, 1H),7.30–7.21(m, 2H),7.13–7.03(m, 2H),6.98(d, J＝7.9 Hz, 1H),4.75(s, 1H),3.83(s, 2H),3.40(s, 3H),3.29(s, 2H),2.98(dd, J＝9.8,5.3 Hz, 2H),1.80(dt, J＝12.4,6.2 Hz, 2H),1.63(d, J＝12.8 Hz, 2H).

Embodiment 6,

Compound A6 1-(4-chloro-2,6-difluorophenyl)-4-((3-(methylamino)phenoxy)methyl)piperidin-4-ol

Compound A5 (23 mg, 0.048 mmol) was dissolved in tetrahydrofuran, and sodium hydroxide (8 mg, 0.19 mmol) aqueous solution (1 mL) was added, and the reaction was carried out at room temperature for 8 h. After TLC monitoring, the reaction was complete, and an appropriate amount of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and the target compound A6 was further separated by silica gel chromatography to obtain 8 mg of white solid with a yield of 43%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.37–7.19(m,2H),6.96(t,J＝8.0Hz,1H),6.13(dd,J＝8.1,2.2Hz,2H),6.09(t,J＝2.2Hz,1H),5.59(q,J＝5.1Hz,1H),4.63(s,1H),3 .70(s,2H),3.38(d,J＝10.9Hz,2H),2.96(d,J＝11.5Hz,2H),2.65(d,J＝5.0Hz, 3H),1.80(m,J＝12.6,4.6Hz,2H),1.58(d,J＝12.9Hz,2H).HR-MS: m/z405.1157 (M+Na) ⁺ .

Embodiment 7,

Compound A7 N-(3-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)phenyl)-1-(difluoro-13-methyl)-12-fluoroformamide

Using 2,2,2-trifluoro-N-(3-hydroxyphenyl)acetamide and 7 as raw materials, referring to the fifth step of the synthesis method of compound A1, compound A7 was obtained as a reddish brown oil with a yield of 18%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ11.21(s, 1H),7.36(t, J＝2.2 Hz, 1H),7.34–7.23(m, 4H),6.84(dd, J＝8.0,2.4 Hz, 1H),4.73(s, 1H),3.79(s, 2H),2.97(d, J＝11.5 Hz, 2H),1.80(m, J＝12.7,4.6 Hz, 2H),1.62(d, J＝12.9 Hz, 2H),1.24(d, J＝7.9 Hz, 2H).HR-MS:m/z 487.0815(M+Na) ⁺ .

Embodiment 8,

Compound A8 N-(3-((1-(4-chloro-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)phenyl)acetamide

Using N-(3-hydroxyphenyl)acetamide and 7 as raw materials, referring to the fifth step of the synthesis method of compound A1, compound A8 was obtained as a white solid with a yield of 18%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.89 (s, 1H), 7.35 (t, J = 2.2Hz, 1H), 7.26 (d, J = 8.9Hz, 2H), 7.17 (t, J = 8.1Hz, 1H), 7.06 (d, J = 8.1Hz, 1H), 6.63 (dd, J = 8.1, 2.4Hz, 1H) ,4.70(s,1H),3.74(s,2H),3.38(d,J＝11.5Hz,2H),2.96(d,J＝11.6Hz,2H),2.03(s,3H),1.79(dt,J＝12.4,6.4Hz,2H),1.60(d,J＝12.9Hz,2H).

Embodiment 9,

Compound A9 1-(4-chloro-2,6-difluorophenyl)-4-((quinoxaline-5-oxy)methyl)piperidin-4-ol

Using quinoxaline-5-ol and 7 as raw materials, referring to the fifth step of the synthesis method of compound A1, compound A9 was obtained as a light yellow solid with a yield of 12%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.95(q,J＝1.9Hz,2H),7.77(t,J＝8.2Hz,1H),7.65(d,J＝8.4Hz,1H),7.34(d,J＝7.8Hz,1H),7.28(d,J＝8.9Hz,2H),4.80(d,J＝7.6H z,1H),4.04(s,2H),3.42(t,J＝10.8Hz,2H),3.01(d,J＝11.5Hz,2H),1.99(m,J＝12.5,4.8Hz,2H),1.70(d,J＝12.8Hz,2H). HR-MS: m/z 428.0948(M+Na) ⁺ .

Embodiment 10

A10 1-(4-chloro-2,6-difluorophenyl)-4-((quinoxaline-2-oxy)methyl)piperidin-4-ol

Using 2-hydroxyquinoxaline and 7 as raw materials, referring to the fifth step of the synthesis method of compound A1, compound A10 was obtained as a light yellow solid with a yield of 15%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.26(s,1H),7.92(d,J＝8.6Hz,1H),7.81(dd,J＝7.9,1.5Hz,1H),7.61(m,J＝8.7,7.2,1.6Hz,1H),7.40–7.34(m,1H),7.29–7.21(m ,2H),4.74(s,1H),4.31(s,2H),3.25(t,J＝11.4Hz,2H),2.93(d,J＝11.5Hz,2H),1.79(m,J＝12.7,4.4Hz,2H),1.61(d,J＝13.1Hz,2H).HR-MS:m/z 428.095 1(M+Na) ⁺ .

Embodiment 11,

Compound A11 1-(4-chloro-2,6-difluorophenyl)-4-((7-methoxy-1,5-naphthyridin-4-yl)oxy)methyl)piperidin-4-ol

Using 7-methoxy-1,5-naphthyridine-4-ol and 7 as raw materials, referring to the fifth step of the synthesis method of compound A1, compound A11 was obtained as a light yellow solid with a yield of 15%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.36(d,J＝1.7Hz,1H),7.87(d,J＝7.8Hz,1H),7.77(d,J＝2.4Hz,1H),7.31–7.20(m,2H),6.13(d,J＝7.8Hz,1H),4.83(s,1H),4.28( s,2H),3.98(d,J＝1.2Hz,3H),3.27(d,J＝3.4Hz,2H),2.93(d,J＝11.7Hz,2H),1.81(dd,J＝13.0,8.6Hz,2H),1.48(d,J＝12.6Hz,2H).HR-MS:m/z 436.1234(M+H ) ⁺ .

Embodiment 12

Compound A12 1-(4-chloro-2,6-difluorophenyl)-4-((3-hydroxyphenyl)(methyl)amino)methyl)piperidin-4-ol

Using 3-hydroxy-N-methylaniline and 7 as raw materials, referring to the fifth step of the synthesis method of compound A1, compound A12 was obtained as a light yellow solid with a yield of 10%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.92 (s, 1H), 7.24 (d, J = 9.0Hz, 2H), 6.90 (t, J = 8.1Hz, 1H), 6.27–6.22 (m, 1H), 6.20 (d, J = 2.3Hz, 1H), 6.02 (dd, J = 7.8, 2.0Hz, 1H) ,4.43(s,1H),3.24(s,2H),2.93(s,3H),2.89(d,J＝12.0Hz,4H),1.71–1.60(m,2H),1.53(d,J＝12.7Hz,2H).HR-MS:405.1158(M+Na) ⁺ .

Embodiment 13

Compound A13 5-((1-(4-chloro-2,6-difluorophenyl)-3-hydroxypiperidin-3-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

The synthesis steps of compound A13 are as follows:

The raw material 8 (100 mg, 0.614 mmol) was dissolved in 6 mL of anhydrous DMF, and anhydrous potassium carbonate (127 mg, 0.921 mmol) was added. After stirring at room temperature for 5 min, the raw material 9 (191 mg, 0.7368 mmol) was slowly added, and the temperature was raised to 80°C for 6 h. The reaction was complete after TLC monitoring. The reaction solution was cooled to room temperature, and an appropriate amount of ice water was added. It was extracted with ethyl acetate three times, and the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain the target compound A13 as a light yellow solid of 42 mg, with a yield of 16%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),7.30–7.21(m,2H),7.05(t,J＝8.1Hz,1H),6.57(d,J＝8.3Hz,1H),6.47(d,J＝7.9Hz,1H),4.78(s,1H),3.98(s,2H),3.22(d,J＝11.8Hz,1 H),3.03(d,J＝5.6Hz,2H),2.94(d,J＝11.8Hz,1H),2.87–2.75(m,2H),2.38(m,J＝7.7,2.2Hz,2H),1.96–1.87(m,1H),1.77(s,1H),1.63–1.46(m,2H). HR-MS:m/z 445.1109(M+Na) ⁺ .

Embodiment 14

Compound A14 1-(4-chloro-2,6-difluorophenyl)-3-((quinoxaline-5-oxy)methyl)piperidin-3-ol

Using quinoxaline-5-ol and 9 as raw materials, referring to the synthesis method of compound A13, compound A14 was obtained as a light yellow solid with a yield of 10%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ8.93(d, J＝1.8 Hz, 1H),8.89(d, J＝1.9 Hz, 1H),7.75(t, J＝8.2 Hz, 1H),7.62(dd, J＝8.4,1.1 Hz, 1H),7.31(dd, J＝7.9,1.2 Hz, 1H),7.23–7.14(m, 2H),4.87(s, 1H),4.26 (s,2H),3.37(d,J＝11.9Hz,1H),3.05(d,J＝4.8Hz,2H),3.02(d,J＝11.6Hz,1H),2.14–2.05(m,1H),1.87–1.77(m,1H),1.68–1.51(m,2H).HR-MS: m/z:428 .0948(M+Na) ⁺ .

Embodiment 15

Compound A15 1-(4-chloro-2,6-difluorophenyl)-3-((quinoxaline-2-oxy)methyl)piperidin-3-ol

Compound A15 was obtained by using 2-hydroxyquinoxaline and 9 as raw materials and referring to the synthesis method of compound A13 as a pale yellow solid with a yield of 13%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ8.59(s,1H),8.01(dd,J＝8.2,1.4Hz,1H),7.83(dd,J＝8.3,1.4Hz,1H),7.75(ddd,J＝8.3,6.9,1.5Hz,1H),7.64(ddd,J＝8.3,7.0,1.5Hz,1H),7.27–7.18(m,2H),4.95(s, 1H),4.55(s,2H),3.26(d,J＝11.8Hz,1H),3.06(s,2H),2.98(d,J＝11.8Hz,1H),1.97(d,J＝12.9Hz,1H),1.83–1.73(m,1H),1.63(td,J＝8.7,4.2Hz,1H) ,1.59–1.48(m,1H).

Example 16

Compound A16 1-(4-chloro-2,6-difluorophenyl)-3-((7-methoxy-1,5-naphthyridin-4-yl)oxy)methyl)piperidin-3-ol

Using 7-methoxy-1,5-naphthyridine-4-ol and 9 as raw materials and referring to the synthesis method of compound A13, compound A16 was obtained as a light yellow solid with a yield of 17%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.36 (d, J = 2.0Hz, 1H), 7.87 (d, J = 7.8Hz, 1H), 7.77 (d, J = 2.5Hz, 1H), 7.31–7.17 (m, 2H), 6.13 (d, J = 7.8Hz, 1H), 4.83 (s, 1H), 4.28 ( s,2H),3.98(d,J＝1.2Hz,3H),3.26(d,J＝12.7Hz,2H),2.93(d,J＝11.7Hz,2H),1.81(dd,J＝13.1,8.6Hz,2H),1.48(d,J＝12.6Hz,2H).

Embodiment 17

Compound A17 5-((4-((4-chloro-2,6-difluorophenyl)amino)-1-hydroxycyclohexyl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

The synthesis steps of compound A17 are as follows:

Intermediate 6 (100 mg, 0.55 mmol) was dissolved in 6 mL of anhydrous DMF, anhydrous potassium carbonate (114 mg, 0.825 mmol) was added, and the raw material 10 (181 mg, 0.66 mmol) was slowly added after stirring at room temperature for 5 min, and the temperature was raised to 80°C for 6 h. The reaction was complete after TLC monitoring, and the reaction solution was cooled to room temperature, and an appropriate amount of ice water was added, and extracted with ethyl acetate three times, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain the target compound A17 as a light yellow solid with a yield of 10%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.98 (s, 1H), 7.36 (d, J = 7.2Hz, 2H), 6.98 (t, J = 9.7Hz, 1H), 6.53 (dd, J = 9.1, 3.8Hz, 1H), 5.76 (s, 1H), 4.46 (s, 1H), 3.67 (s, 2H), 3. 57(s,2H),2.88(t,J＝7.6Hz,2H),2.44(t,J＝7.7Hz,2H),2.43–2.31(m,2H),1.66(t,J＝12.4Hz,2H),1.51(d,J＝12.9Hz,2H).HR-MS:m/z 455.1342(M+H) ⁺ .

Embodiment 18

Compound A18 4-((4-((4-chloro-2,6-difluorophenyl)amino)-1-hydroxycyclohexyl)methoxy)quinolin-2(1H)-one

Compound A18 was obtained by using 4-hydroxyquinolin-2(1H)-one and 10 as raw materials and referring to the synthesis method of compound A17. The compound A18 was a pale yellow solid with a yield of 12%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ7.84–7.79 (m, 1H), 7.25 (t, J＝7.8 Hz, 1H), 7.16 (d, J＝8.0 Hz, 2H), 6.76 (d, J＝8.4 Hz, 1H), 6.62 (s, 2H), 6.54 (t, J＝7.5 Hz, 1H), 4.70 (d, J＝7.9 Hz, 1H), 4.56 (s, 1H), 3.99 (s, 2H), 3.76 (m, 1H), 1.65 (m, 5H), 1.43 (m, 2H).

Embodiment 19

Compound A19 4.-((4-chloro-2,6-difluorophenyl)amino)-1-((quinoxaline-5-oxy)methyl)cyclohexane-1-ol

Using quinoxaline-5-ol and 10 as raw materials, compound A19 was obtained as a pale yellow solid with a yield of 11% by referring to the synthetic method of compound A17. ¹ H NMR (400 MHz, DMSO-d ₆ )δ9.00–8.86 (m, 2H), 7.76 (t, J＝8.1 Hz, 1H), 7.64 (d, J＝8.4 Hz, 1H), 7.30 (d, J＝7.8 Hz, 1H), 7.18 (d, J＝8.1 Hz, 2H), 4.78–4.68 (m, 1H), 4.58 (s, 1H), 3.97 (s, 2H), 3.60–3.37 (m, 1H), 1.71 (p, J＝12.5, 12.0 Hz, 7H). HR-MS: m/z 442.1101 (M+Na) ⁺ .

Embodiment 20,

Compound A20 4-((4-chloro-2,6-difluorophenyl)amino)-1-((quinoxaline-2-oxy)methyl)cyclohexane-1-ol

Using 2-hydroxyquinoxaline and 10 as raw materials and referring to the synthesis method of compound A17, compound A20 was obtained as a light yellow solid with a yield of 10%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.61 (s, 1H), 8.01 (d, J = 8.2Hz, 1H), 7.84 (dd, J = 8.5, 1.5Hz, 1H), 7.79–7.73 (m, 1H), 7.64 (m, J = 8.4, 6.9, 1.5Hz, 1H), 7.23–7.13 (m,2H),4.77–4.65(m,1H),4.59(s,1H),4.25(s,2H),1.71(t,J＝11.8Hz,6H),1.56(d,J＝12.5Hz,2H).HR-MS: m/z 442.1180(M+Na) ⁺ .

Embodiment 21,

Compound B1 8-Fluoro-5-((4-hydroxypiperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

The synthesis steps of compound B1 are as follows:

first step:

Dissolve intermediate 6 (0.1 g, 0.55 mmol) and anhydrous potassium carbonate (0.092 g, 0.66 mmol) in 4 mL DMF, stir for half an hour, add raw material 11 (0.14 g, 0.66 mmol), and react at 80°C for 6-8 hours before the reaction is complete. Stop the reaction, add water to the reaction solution, and extract with ethyl acetate. Dry, dry and concentrate the organic phase under reduced pressure, and separate by silica gel column chromatography. Intermediate 12 is obtained, 100 mg, with a yield of 46%.

Step 2:

Dissolve intermediate 12 (50 mg, 0.275 mmol) in methanol, add 5 mL of concentrated hydrochloric acid, and react at room temperature for 2 h until the reaction is complete. Distill the reaction solution under reduced pressure to dryness, add a small amount of alkaline water, and extract with ethyl acetate. Dry, concentrate the organic phase under reduced pressure, and separate by silica gel column chromatography to obtain compound B1. A7 (compound F1) is obtained, 36 mg of white solid, with a yield of 90%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.03 (s, 1H), 7.07–6.96 (m, 1H), 6.58 (dd, J = 9.1, 3.7Hz, 1H), 5.17 (s, 1H), 3.77 (s, 2H), 3.18 (t, J = 4.7Hz, 2H), 3.15–3.07 (m, 2H), 2.91 (t, J＝7.6Hz, 2H), 2.48 (s, 2H), 1.92 (m, J＝13.6, 4.7Hz, 2H), 1.69 (d, J＝13.9Hz, 2H). HR-MS: m/z295.1459 (M+H) ⁺ .

Embodiment 22,

Compound B2 5-((1-(2,6-difluoro-4-nitrophenyl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Compound B1 (300 mg, 1.02 mmol) was dissolved in 10 mL of anhydrous DMF, DIEA (337 μL, 2.04 mmol) was added, 1,2,3-trifluoro-5-nitrobenzene (143 μL, 1.224 mmol) was slowly added after stirring at room temperature for 5 min, the temperature was raised to 55°C, and the reaction was continued for 10 h. TLC monitored the reaction to be complete, the reaction solution was cooled to room temperature, an appropriate amount of ice water was added, and it was extracted three times with ethyl acetate, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, the filtrate was concentrated to dryness under reduced pressure, and the target compound B2 was further separated by silica gel chromatography column, 280 mg of yellow solid, and the yield was 60.8%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.00 (s, 1H), 8.06–7.72 (m, 2H), 7.01 (t, J = 9.7Hz, 1H), 6.59 (dd, J = 9.1, 3.8Hz, 1H), 4.82 (s, 1H), 3.78 (s, 2H), 3.51 (t, J = 12. 0Hz,2H),3.34(s,2H),2.91(t,J＝7.5Hz,2H),2.50–2.41(m,2H),1.83(m,J＝12.7,4.5Hz,2H),1.67(d,J＝13.0Hz,2H). HR-MS: m/z 474.1257(M+Na) ⁺ .

Embodiment 23,

Compound B3 5-((1-(4-amino-2,6-difluorophenyl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Compound B2 (75 mg, 0.17 mmol) was dissolved in 10 mL of methanol, 10% palladium carbon (19 mg, 0.017 mmol) was added, the mixture was filtered under reduced pressure, and replaced with hydrogen three times. The reaction system was reacted at room temperature for 4 h under a hydrogen environment. TLC monitored the reaction to be complete, the reaction solution was filtered through diatomaceous earth to remove palladium carbon, the filter cake was rinsed with methanol three times, the filtrate was concentrated to dryness under reduced pressure, and finally separated by silica gel chromatography to obtain compound B3, 50 mg of white solid, with a yield of 71%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.00(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.8Hz,1H),6.20–6.06(m,2H),5.39(s,2H),4.56(s,1H),3.75(s,2H),3.28(d,J＝11.5Hz ,2H),2.92(t,J＝7.6Hz,2H),2.77–2.67(m,2H),2.51–2.42(m,2H),1.75(dt,J＝12.1,6.3Hz,2H),1.59(d,J＝12.7Hz,2H).HR-MS:m/z 444.1499(M+Na) ⁺ .

Embodiment 24,

Compound B4 N-(3,5-difluoro-4-(4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-4-hydroxypiperidin-1-yl)phenyl)acetamide

Compound B3 (15 mg, 0.036 mmol) was dissolved in 5 mL of dichloromethane, triethylamine (10 μL, 0.07 mmol) was added, and the mixture was stirred at 0°C for 2 min. Acetyl chloride (4 μL, 0.04 mmol) was slowly added dropwise, and the mixture was stirred at 0°C for 30 min, and then the mixture was naturally warmed to room temperature and the reaction was continued for 2 h. The reaction was complete after TLC monitoring, and the reaction solution was concentrated under reduced pressure and dried, and then separated by silica gel chromatography to obtain compound B4, 19 mg of white solid, with a yield of 33%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.13(s,1H),10.00(s,1H),7.43–7.18(m,2H),7.00(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.7Hz,1H),4.65(s,1H),3.76(s,2H), 3.37(d,J＝12.2Hz,2H),2.97–2.83(m,4H),2.49–2.44(m,2H),2.03(s,3H),1.80(m,J＝12.5,4.5Hz,2H),1.62(d,J＝12.7Hz,2H).

Embodiment 25

Compound B5 N-(3,5-difluoro-4-(4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-4-hydroxypiperidin-1-yl)phenyl)-N-(methylsulfonyl)methanesulfonamide

Compound B5 was obtained with compound B3 and methanesulfonyl chloride as raw materials and referring to the synthesis method of compound B4 as a white solid with a yield of 63%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.00 (s, 1H), 7.38 (d, J = 9.4Hz, 2H), 7.01 (t, J = 9.7Hz, 1H), 6.59 (dd, J = 9.1, 3.7Hz, 1H), 4.74 (s, 1H), 3.78 (s, 2H), 3.54 (s, 6H), 3 .43(t,J＝11.7Hz,2H),3.11(d,J＝11.7Hz,2H),2.92(t,J＝7.7Hz,2H),2.46(d,J＝10.9Hz,2H),1.90–1.76(m,2H),1.65(d,J＝12.9Hz,2H).

Embodiment 26

Compound B6 5-((1-(3,5-difluoropyridin-4-yl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using compound B1 and 3,4,5-trifluoropyridine as raw materials and referring to the synthesis method of compound B2, compound B6 was obtained as a white solid with a yield of 40%. ¹ H NMR (400MHz, Chloroform-d) δ8.18–8.08(m,2H),7.62(s,1H),6.93(t,J＝9.4Hz,1H),6.48(dd,J＝9.1,3.9Hz,1H),3.85(s,2H),3.66–3.53(m,2H),3.45(d,J＝ 12.9Hz,2H),3.01(t,J＝7.7Hz,2H),2.66(dd,J＝8.4,6.9Hz,2H),2.14(s,1H),1.95–1.80(m,4H).HR-MS: m/z 430.1355(M+Na) ⁺ .

Embodiment 27

Compound B7 3,5-difluoro-4-(4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-4-hydroxypiperidin-1-yl)benzoic acid methyl ester

Using compound B1 and methyl 3,4,5-trifluorobenzoate as raw materials and referring to the synthesis method of compound B2, compound B7 was obtained as a white solid with a yield of 20%. ¹ H NMR (400MHz, Chloroform-d) δ7.58(s,1H),7.55–7.47(m,2H),6.93(t,J＝9.4Hz,1H),6.49(dd,J＝9.1,3.9Hz,1H),3.89(s,3H),3.85(s,2H),3.56(t,J＝11.7 Hz,2H),3.29(d,J＝12.5Hz,2H),3.02(t,J＝7.7Hz,2H),2.66(dd,J＝8.4,6.9Hz,2H),2.14(s,1H),1.95–1.78(m,4H).HR-MS: m/z 487.1452(M+Na) ⁺ .

Embodiment 28

Compound B8 8-Fluoro-5-((4-hydroxy-1-isobutylpiperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

Compound B1 (100 mg, 0.34 mmol) was dissolved in 4 mL of anhydrous DMF, DIEA (222 mg, 0.68 mmol) was added, and after stirring at room temperature for 5 min, isobutyl bromide (45 μL, 0.408 mmol) was added, and the temperature was immediately raised to 65°C, and the reaction was continued for 4 h. TLC monitored the reaction to be complete, the reaction solution was cooled to room temperature, an appropriate amount of ice water was added, and the mixture was extracted three times with ethyl acetate, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and the target compound B8 was further separated by silica gel chromatography column, as a white solid of 21 mg, with a yield of 18%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.00 (s, 1H), 7.01 (t, J = 9.7Hz, 1H), 6.59 (dd, J = 9.1, 3.7Hz, 1H), 4.74 (s, 1H), 3.78 (s, 2H), 3.54 (s, 6H), 3.11 (d, J = 11.7Hz, 2H) ,2.92(t,J＝7.7Hz,2H),2.45(s,2H),1.88–1.76(m,1H),1.65(d,J＝12.9Hz,2H),1.25(d,J＝8.8Hz,6H).

Embodiment 29,

Compound B9 8-Fluoro-5-((4-hydroxy-1-(pentan-3-yl)piperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

Compound B9 was obtained by using compound B1 and 3-bromopentane as raw materials and referring to the synthesis method of compound B8. The white solid compound B9 had a yield of 19%. ¹ H NMR (400 MHz, Chloroform-d) δ7.57 (s, 1H), 6.92 (t, J = 9.4 Hz, 1H), 6.46 (dd, J = 9.1, 3.9 Hz, 1H), 5.30 (s, 1H), 4.66 (p, J = 6.1 Hz, 1H), 4.01 (d, J = 13.2 Hz, 2H), 3.80 (s, 2H), 3 .27(t,J＝12.8Hz,2H),3.04–2.88(m,2H),2.64(dd,J＝8.4,7.0Hz,2H),2.04(s,1H),1.74(d,J＝13.3Hz,2H),1.62–1.50(m,4H),1.36–1.19(m,2H),0.9 0(t,J＝7.4Hz,6H).

Embodiment 30

Compound B10 5-((1-cyclopentyl-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using compound B1 and bromocyclopentane as raw materials and referring to the synthesis method of compound B8, compound B10 was obtained as a white solid with a yield of 21%. ¹ H NMR (400MHz, Chloroform-d) δ7.61(s,1H),6.84(t,J＝9.4Hz,1H),6.38(dd,J＝9.1,3.9Hz,1H),5.05(tt,J＝6.1,2.7Hz,1H),3.89(s,2H),3.72(s,2H),3.17(t, J＝12.4Hz,2H),2.92(t,J＝7.6Hz,2H),2.57(t,J＝7.7Hz,2H),2.16–1.91(m,1H),1.78(m,J＝16.1,5.3Hz,2H),1.71–1.48(m,8H),1.20(d,J＝11.7Hz,2H).

Embodiment 31,

Compound B11 5-((1-cyclohexyl-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using compound B1 and bromocyclohexane as raw materials and referring to the synthesis method of compound B8, compound B11 was obtained as a white solid with a yield of 18%. ¹ H NMR (400MHz, Chloroform-d) δ7.57 (s, 1H), 6.92 (t, J = 9.4Hz, 1H), 6.46 (dd, J = 9.1, 3.9Hz, 1H), 5.30 (s, 1H), 4.66 (h, J = 5.8, 5.4Hz, 1H), 4.01 (d, J = 13.1Hz, 2H) ,3.80(s,2H),3.27(t,J＝12.8Hz,2H),2.99(t,J＝7.7Hz,2H),2.64(dd,J＝8.4,7.0Hz,2H),2.04(s,1H),1.74(d,J＝13.3Hz,2H),1.61–1.51(m,4H),0.90( t,J＝7.4Hz,8H).

Embodiment 32

Compound B12 8-Fluoro-5-((4-hydroxy-1-(2-(tetrahydro-2H-pyran-4-yl)ethyl)piperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

Compound B1 and 4-(2-bromoethyl)tetrahydro-2H-pyran were used as raw materials, and the synthesis method of compound B8 was referred to to obtain compound B12 as a white solid with a yield of 16%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),6.99(t,J＝9.7Hz,1H),6.55(dd,J＝9.1,3.7Hz,1H),4.54(s,1H),3.81(dd,J＝10.9,4.2Hz,2H),3.70(s,2H),3.30–3.20(m,3H),2.90(t,J ＝7.6Hz,2H),2.76–2.53(m,4H),2.47–2.29(m,5H),1.70(d,J＝12.9Hz,2H),1.63–1.50(m,3H),1.40(s,1H),1.24(s,1H),1.15(tt,J＝12.1,6.3Hz,2H ).HR-MS:m/z 407.2339(M+H) ⁺ .

Embodiment 33,

Compound B13 8-Fluoro-5-((4-hydroxy-1-(2-morpholinoethyl)piperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

Compound B1 and 4-( ² -bromoethyl)morpholine were used as raw materials, and the synthesis method of compound B8 was referred to to obtain compound B13 as a white solid with a yield of 12%. NMR (400MHz, Chloroform-d) δ7.61(s,1H),6.92(t,J＝9.4Hz,1H),6.45(dd,J＝9.1,3.9Hz,1H),3.87(s,2H),3.78–3.68(m,6H),3.67–3.62(m,1H),3.48(d,J＝11. 2Hz,2H),3.15–3.06(m,1H),2.99(dd,J＝8.5,7.0Hz,2H),2.86(t,J＝6.2Hz,2H),2.64(dd,J＝8.5,6.9Hz,2H),2.61–2.52(m,4H),2.38(s,2H),1.91(d,J＝1 4.2Hz,2H).HR-MS:m/z 408.2292(M+H) ⁺ .

Embodiment 34,

Compound B14 5-((1-(4-chloro-2,6-difluorobenzyl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using compound B1 and 4-chloro-2,6-difluorobenzyl bromide as raw materials and referring to the synthesis method of compound B8, compound B14 was obtained as a white solid with a yield of 7%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.98(s,1H),7.23–7.11(m,2H),6.99(t,J＝9.7Hz,1H),6.54(dd,J＝9.1,3.8Hz,1H),4.47(s,1H),3.68(s,2H),2.90(t,J＝7.6Hz ,2H),2.57(d,J＝4.1Hz,1H),2.46(dd,J＝8.5,6.9Hz,2H),1.67(t,J＝8.4Hz,6H),1.50(q,J＝12.2,11.1Hz,1H).HR-MS: m/z 477.1189(M+Na) ⁺ .

Embodiment 35

Compound B15 5-((1-(4-chloro-2,6-difluorobenzoyl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

4-Chloro-2,6-difluorobenzoic acid (25 μL, 0.21 mmol) was dissolved in 8 mL of anhydrous DMF, and DIEA (75 μL, 0.42 mmol) and HATU (88 mg, 0.231 mmol) were added in sequence. The mixture was stirred at room temperature for 10 min, and then compound B1 (74 mg, 0.252 mmol) was added. The reaction was continued at room temperature for 2 h. TLC monitored the reaction to be complete, and an appropriate amount of water was added. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and the target compound B15 was further separated by silica gel chromatography column as a white solid of 62 mg with a yield of 64%. ¹ H NMR(400MHz,Chloroform-d)δ7.62(s,1H),7.01(m,J＝7.2,6.2,2.2Hz,2H),6.92(t,J＝9.4Hz,1H),6.45(dd,J＝9.1,3.9Hz,1H),4.66(dt,J＝13.4,3.3Hz,1H),3.93– 3.71(m,2H),3.56(m,J＝12.9,3.3H z,1H),3.39(d,J＝13.5Hz,1H),3.30(m,J＝12.9,3.3Hz,1H),2.98(t,J＝7.7Hz,2H),2.64(t,J＝7.7Hz,2H),2.19(s,1H),1.89(dd,J＝13.7,2.9Hz,1H),1.82 –1.73(m,2H),1.72–1.65(m,1H).HR-MS:m/z 491.0961(M+Na) ⁺ .

Embodiment 36

Compound B16 5-((1-((3r, 5r, 7r)-adamantane-1-carbonyl)-4-hydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using adamantanecarboxylic acid and compound B1 as raw materials and referring to the synthesis method of compound B15, compound B16 was obtained as a white solid with a yield of 58%. ¹ H NMR (400MHz, Chloroform-d) δ7.67 (s, 1H), 6.92 (t, J = 9.4Hz, 1H), 6.45 (dd, J = 9.1, 3.9Hz, 1H), 4.44–4.25 (m, 2H), 3.79 (s, 2H), 3.29 (t, J = 12.7Hz, 2H), 2.99 ( dd,J＝8.4,6.9Hz,2H),2.65(dd,J＝8.5,6.9Hz,2H),2.08–1.98(m,9H),1.98–1.90(m,1H),1.78(d,J＝13.2Hz,5H),1.65(dt,J＝13.1,6.6Hz,4H).

Embodiment 37

Compound C1 N-(5-(((3R, 4R)-1-(4-chloro-2,6-difluorophenyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-2-fluorophenyl)-1-(difluoro-13-methyl)-12-fluoroformamide

The synthetic route of compound C1 is as follows:

first step:

The raw material 13 (48 g, 195.40 mmol), (S)-5-(pyrrolidin-2-yl)-1H-tetrazole (5.44 g, 39.08 mmol) and anhydrous sodium acetate (3.21 g, 39.08 mmol) were dissolved in 200 mL of anhydrous DMF, cooled to -40 °C, and a 3/4 solution of nitrosobenzene (20.93 g, 195.40 mmol) in DMF (300 mL) was added under N ₂ protection. The reaction system was reacted at -30 °C for 4 h under N ₂ protection. Then, the remaining 1/4 nitrosobenzene was added, and the reaction was continued for 6 h under N ₂ protection at -30 to -40 °C. A saturated aqueous ammonium chloride solution was added, and the mixture was extracted twice with methyl tert-butyl ether. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain a reaction intermediate. The reaction intermediate was dissolved in methanol (240 mL), the reaction system was cooled to -10 ° C, anhydrous copper sulfate (15.59 g, 97.70 mmol) was added, and the reaction was continued for 2 h. TLC monitored the reaction to be complete, and an appropriate amount of water was added to dilute the reaction solution, extracted three times with methyl tert-butyl ether, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure and further separated by silica gel chromatography to obtain intermediate 14, 13.1 g of brown oil, with a yield of 26%.

Step 2:

Intermediate 14 (12 g, 45.86 mmol) and trimethyl sulfoxide iodide (13.12 g, 59.62 mmol) were dissolved in dimethyl sulfoxide (145 mL), and sodium tert-butoxide (5.73 g, 59.62 mmol) was added under _N2 protection, and stirred at room temperature for 20 min. TLC monitored the reaction to be complete, and an appropriate amount of water was added, extracted three times with ethyl acetate, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain a racemic compound, and then separated by chiral column to obtain intermediate 15, 3.6 g yellow solid, yield 29%.

Step 3:

The raw material 16 (60 mg, 0.27 mmol) was dissolved in 5 mL of anhydrous DMF, anhydrous potassium carbonate (75 mg, 0.54 mmol) was added, and the intermediate 15 (82 mg, 0.297 mmol) was slowly added after stirring at room temperature for 5 min, and the temperature was raised to 80°C for 6 h. The reaction was complete after TLC monitoring, and the reaction solution was cooled to room temperature, and an appropriate amount of ice water was added, extracted with ethyl acetate three times, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain the target compound C1, 10 mg of brown oil, with a yield of 7%. ¹ H NMR(400MHz, DMSO-d ₆ )δ8.93(s,1H),7.31–7.21(m,2H),6.77(dd,J＝11.8,8.6Hz,1H),6.23(dd,J＝7.7,2.8Hz,1H),5.88(dt,J＝8.6,3.2Hz,1H),5.07–4.83 (m,2H),4.36(s,1H),3.64–3.49(m,1H),3.31–3.19(m,2H),3.18–3.09(m,2H),2.94(dd,J＝11.0,5.0Hz,1H),2.84(d,J＝11.6Hz,1H).HR-MS:m/z425.0871 (MC ₂ F ₃ O) ⁺ .

Embodiment 38

Compound C2 N-(3-(((3R, 4R)-1-(4-chloro-2,6-difluorophenyl)-3,4-dihydroxypiperidin-4-yl)methoxy)phenyl)-1-(difluoro-13-methyl)-12-fluoroformamide

Using 2,2,2-trifluoro-N-(3-hydroxyphenyl)acetamide and intermediate 15 as raw materials, referring to the third step synthesis method of compound C1, compound C2 was obtained as a reddish brown oil with a yield of 8%. ¹ H NMR (400MHz, DMSO-d6) δ8.87 (s, 1H), 7.25 (d, J = 8.8 Hz, 2H), 6.81 (t, J = 8.0 Hz, 1H), 6.13 (d, J = 2.1 Hz, 1H), 6.08 (d, J = 2.3 Hz, 1H), 5.94 (dd, J = 7.9, 2.1 Hz, 1H), 5.22 (t, J = 6 .0Hz,1H),4.74(s,1H),4.23(s,1H),3.58(s,1H),3.31–3.21(m,1H),3.16(t,J＝10.0Hz,2H),3.01(dd,J＝13.0,5.2Hz,1H),2.93(d,J＝7.9Hz,1H),2.84 (d,J＝11.0Hz,1H).

Embodiment 39,

Compound C3 5-((3R, 4R)-1-(4-chloro-2,6-difluorophenyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-3,4-dihydroquinolin-2(1H)-one

Using 5-hydroxy-3,4-dihydroquinolin-2(1H)-one and intermediate 15 as raw materials, referring to the third step synthesis method of compound C1, compound C3 was obtained as a white solid with a yield of 20%. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.02(s,1H),7.31–7.23(m,2H),7.08(t,J＝8.1Hz,1H),6.59(d,J＝8.3Hz,1H),6.49(d,J＝8.0Hz,1H),4.86(d,J＝6.4Hz,1H),4.51(s,1H),4.04(d,J＝8.8Hz,1H),3.75(dt,J＝11.0,5.6Hz,1 H),3.68(d,J＝8.8Hz,1H),3.39–3.31(m,1H),3.21(t,J＝10.7Hz,1H),2.97(dd,J＝11.0,5.1Hz,1H),2.92–2.80(m,3H),2.43(t,J＝7.7Hz,2H),1.92(m,J＝ 13.0,4.8Hz,1H),1.74–1.59(m,1H).,

Embodiment 40

Compound C4 7-((3R, 4R)-1-(4-chloro-2,6-difluorophenyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-4-fluoro-1,1a,3,7b-tetrahydro-2H-cyclopropane[c]quinolin-2-one

Using 4-fluoro-7-hydroxy-1,1a,3,7b-tetrahydro-2H-cyclopropane[c]quinolin-2-one and intermediate 15 as raw materials, referring to the third step of the synthesis method of compound C1, compound C4 was obtained as a light yellow solid with a yield of 20%. ¹ H NMR (400MHz, DMSO-d ₆ )δ9.76(s,1H),7.31–7.23(m,2H),7.02–6.94(m,1H),6.57(m,J＝9.3,5.9,3.8Hz,1H),4.87(dd,J＝11.7,6.5Hz,1H),4.54(d,J＝4.5Hz,1H),4.07(t,J＝8. 3Hz,1H),3.81–3.69(m,2H),3.35(d,J＝11.6Hz,1H),3.26–3.17(m,1H),2. 96(dd,J＝11.0,5.1Hz,1H),2.90(d,J＝8.3Hz,1H),2.78–2.67(m,1H),2.04(dt,J＝8.3,4.4Hz,1H),1.99–1.86(m,1H),1.73(dd,J＝19.4,13.5Hz,1H),1.6 5(m,J＝9.2,4.3Hz,1H),0.57(p,J＝4.5Hz,1H).HR-MS:m/z491.0961(M+Na) ⁺ .

Embodiment 41,

Compound D1 tert-butyl (4R)-4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-3,4-dihydroxypiperidine-1-carboxylate

The synthetic route of compound D1 is as follows:

first step:

The raw material 17 (1 g, 4.69 mmol) was dissolved in toluene (15 mL), triethylamine (716 μL, 5.16 mmol) was added at room temperature, and then thionyl chloride (375 μL, 5.16 mmol) was slowly added dropwise, and the mixture was stirred for 5 min at room temperature and then heated to 40°C for 30 min. The reaction was complete when monitored by TLC, and ice water was added to the reaction solution, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain the crude intermediate 18, which was directly used in the next step without further purification.

Step 2:

Intermediate 6 (412 mg, 2.265 mmol) was dissolved in anhydrous DMF (15 mL), anhydrous potassium carbonate (785 mg, 5.67 mmol) was added, and intermediate 18 (4.69 mmol) was added after stirring at room temperature for 5 min, and the temperature was immediately raised to 80°C for 4 h. TLC monitored the reaction to be complete, the reaction solution was cooled to room temperature, an appropriate amount of ice water was added, and it was extracted with ethyl acetate three times, the organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain intermediate 19, 731 mg of white solid, with a yield of 86%.

Step 3:

AD-mixβ (935 mg, 1.2 mmol) and methanesulfonamide (77 mg, 0.8 mmol) were added to a mixed solution of water (10 mL) and tert-butanol (10 mL) in sequence, stirred at room temperature for 10 min, then cooled to 0°C, and then intermediate 19 (300 mg, 0.8 mmol) was added, and the mixture was reacted at 0°C for 15 h. After TLC monitoring, the reaction was complete, anhydrous sodium sulfite (504 mg, 4 mmol) was added, stirred at room temperature for 30 min, diluted with water, extracted three times with ethyl acetate, the organic phases were combined, washed with 1 M NaOH, washed with water, dried over anhydrous sodium sulfate, and filtered, the filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain compound D1, 189 mg of white solid, with a yield of 58%. ² .63(q,J=8.0,7.1Hz,3H),1.66(s,2H),1.47(s,9H).

Embodiment 42,

Compound D2 5-((3R, 4R)-1-(2,6-difluoro-4-nitrophenyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

The synthesis steps of compound D2 are as follows:

first step:

Compound D1 (335 mg, 0.82 mmol) was dissolved in dichloromethane (8 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 4 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated to dryness under reduced pressure at 40 °C, and then dissolved in an appropriate amount of dichloromethane and concentrated to dryness under reduced pressure. The operation was repeated 3 times to obtain a crude product intermediate 20, which was directly used in the next step without further purification.

Step 2:

The intermediate 20 (300 mg, 0.97 mmol) was dissolved in 10 mL of anhydrous DMF, and DIEA (344 μL, 1.94 mmol) was added. After stirring at room temperature for 5 min, 1,2,3-trifluoro-5-nitrobenzene (raw material 21) (136 μL, 1.164 mmol) was slowly added, and the temperature was raised to 55°C, and the reaction was continued for 10 h. TLC monitored the reaction to be complete, the reaction solution was cooled to room temperature, an appropriate amount of ice water was added, and it was extracted three times with ethyl acetate. The organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain the target compound D2, 220 mg of yellow solid, with a yield of 50%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.01(s,1H),8.06–7.92(m,2H),7.02(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.7Hz,1H),5.01(d,J＝6.3Hz,1H),4.66(s,1H),4.04(d,J＝8.8Hz,1H),3.76(d t,J＝8.8,6.5Hz, 1H),3.70(d,J＝8.8Hz,1H),3.45(t,J＝12.3Hz,1H),3.26(d,J＝17.0Hz,2H),2.87(m,J＝7.6,4.6Hz,2H),2.50–2.42(m,3H),1.94(m,J＝13.2,4.8Hz,1H), 1.78–1.69(m,1H).

Embodiment 43,

Compound D3 5-((3R, 4R)-1-(4-amino-2,6-difluorophenyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Compound D3 was obtained by using compound D2 as raw material and referring to the synthesis method of compound B3. The yield was 60%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),7.01(t,J＝9.7Hz,1H),6.57(dd,J＝9.1,3.7Hz,1H),6.14(d,J＝11.5Hz,2H),5.41(s,2H),4.69(d,J＝6.5Hz,1H),4.36(s,1H),3.99(d,J＝8.7Hz,1H),3.68( m,J＝9.9,9.0,4.8Hz,2H),3.31–3.21(m,1H),3.15(t,J＝10.4Hz,1H),2.93–2.86(m,1H),2.71(dd,J＝10.4,5.1Hz,1H),2.47(d,J＝7.8Hz,4H),1.85(dt,J＝ 12.8,6.9Hz,1H).

Embodiment 44,

Compound D4 N-(3,5-difluoro-4-((3R, 4R)-4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-3,4-dihydroxypiperidin-1-yl)phenyl)acetamide

Compound D3 was used as the raw material, and the synthesis method of compound B4 was referred to obtain compound D4, a white solid, with a yield of 65%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.13(s,1H),10.01(s,1H),7.24(d,J＝11.3Hz,2H),7.02(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.8Hz,1H),4.80(d,J＝6.5Hz,1H),4.46(s,1H),4.01(d,J ＝8.8Hz,1H),3.78–3.65(m,2H),3.20(t,J＝10.5Hz,1H),2.89(dd,J＝7.5,5.1Hz,2H),2.80(d,J＝11.0Hz,1H),2.47(d,J＝7.9Hz,4H),2.03(s,3H),1.95–1 .85(m,1H).

Embodiment 45

Compound D5 5-((3R, 4R)-1-(3,5-difluoropyridin-4-yl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 3,4,5-trifluoropyridine as raw materials, referring to the second step synthesis method of compound D2, compound D5 was obtained as an off-white solid with a yield of 25%. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.01(s,1H),8.26(s,2H),7.02(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.7Hz,1H),5.76(s,1H),4.99(d,J＝6.3Hz,1H),4.64(s,1H),4.03(d,J＝8.8Hz,1H),3.76(dd,J＝10.6, 5.4Hz,1H),3.69(d,J＝8.8Hz,1H),3.41(t,J＝12.3Hz,1H),3.27(d,J＝11.0Hz,1H),2.87(m,J＝7.6,4.6Hz,2H),2.46(t,J＝7.7Hz,2H),1.92(m,J＝13.0,4. 7Hz,1H).HR-MS:m/z 424.1486(M+H) ⁺ .

Embodiment 46

Compound D6 3,5-difluoro-4-((3R, 4R)-4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-3,4-dihydroxypiperidin-1-yl)benzoic acid methyl ester

Using intermediate 20 and methyl 3,4,5-trifluorobenzoate as raw materials, referring to the second step synthesis method of compound D2, compound D6 was obtained as a white solid with a yield of 32%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.01 (s, 1H), 7.52 (d, J = 9.2Hz, 2H), 7.02 (t, J = 9.8Hz, 1H), 6.68–6.49 (m, 1H), 4.94 (d, J = 6.2Hz, 1H), 4.59 (s, 1H), 4.03 (d, J = 8. 8Hz,1H),3.83(s,3H),3.75(s,1H),3.68(d,J＝8.6Hz,1H),3.21–3.09(m,1H),2.87(d,J＝7.5Hz,2H),2.50–2.41(m,4H),1.93(t,J＝12.8Hz,1H).

Embodiment 47

Compound D7 3,5-difluoro-4-((3R, 4R)-4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-3,4-dihydroxypiperidin-1-yl)-N-methylbenzamide

The synthesis steps of compound D7 are as follows:

first step:

Compound D6 (200 mg, 0.42 mmol) was dissolved in methanol (10 mL), and NaOH (84 mg, 2.1 mmol) was weighed and dissolved in 1 mL of water, then added dropwise to the reaction solution, and the temperature was raised to 60°C for 3 h. The reaction was complete as monitored by TLC, and the reaction solution was cooled to room temperature, acidified with 2M hydrochloric acid to a pH of 5-6, and concentrated to dryness under reduced pressure. Then, the solution was dissolved with an appropriate amount of a mixed solvent of dichloromethane/methanol (10/1), filtered to remove the generated NaCl, and the filtrate was concentrated under reduced pressure and dried to obtain a crude intermediate 21, which was used directly in the next step without further purification.

Step 2:

The intermediate 21 (0.1 mmol) was dissolved in DMF, and DIEA (71 μL, 0.4 mmol), HATU (42 mg. 0.11 mmol) and methylamine hydrochloride (10 mg, 0.12 mmol) were added in sequence under stirring at room temperature, and the reaction was carried out at room temperature for 2 h. After TLC monitoring, the reaction was complete, and an appropriate amount of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with water, and then washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure, and further separated by silica gel chromatography to obtain the target compound D7, 15 mg of white solid, with a yield of 31%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.01(s,1H),8.45(q,J＝4.5Hz,1H),7.54–7.45(m,2H),7.02(t,J＝9.7Hz,1H),6.58(dd,J＝9.1,3.7Hz,1H),4.90(s,1H),4.56(s,1H),4.03(d,J＝8.8 Hz,1H),3.75(dd,J＝10.4,5.1Hz,1H),3.68(d,J＝8.8H z,1H),3.24(t,J＝11.1Hz,1H),3.17(s,2H),3.08(dd,J＝28.7,8.6Hz,1H),2.89(m,J＝7.9,7.5,4.8Hz,2H),2.76(d,J＝4.4Hz,2H),2.46(d,J＝7.7Hz,2H),1 .93(m,J＝13.1,4.8Hz,1H),1.70(d,J＝13.7Hz,1H).

Embodiment 48

Compound D8 3,5-difluoro-4-((3R, 4R)-4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-3,4-dihydroxypiperidin-1-yl)-N-isopropylbenzamide

Using intermediate 21 and isopropylamine as raw materials, referring to the second step synthesis method of compound D7, compound D8 was obtained as a white solid with a yield of 42%. ¹ H NMR (400MHz, Chloroform-d) δ8.03 (s, 2H), 7.58 (s, 1H), 7.30–7.27 (m, 1H), 6.93 (t, J = 9.3Hz, 1H), 6.52 (dd, J = 9.0, 3.9Hz, 1H), 5.83 (s, 1H), 4.25 (d, J = 7.6Hz, 1H),4.08–3.95(m,3H),3.54–3.40(m,1H),3.34(d,J＝8.6Hz,2H),3.16(d,J＝12.1Hz,1H),2.97(s,4H),2.89(s,4H),2.69–2.61(m,2H),1.97(d,J＝11 .9Hz,2H).

Embodiment 49

Compound D9 N-cyclopropyl-3,5-difluoro-4-((3R,4R)-4-((8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)methyl)-3,4-dihydroxypiperidin-1-yl)benzamide

Using intermediate 21 and cyclopropylamine as raw materials, compound D9 was obtained as a white solid with a yield of 55% by referring to the second step synthesis method of compound D7. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.01(s, 1H),8.40(d, J＝4.2Hz, 1H),7.50(d, J＝10.0Hz, 2H),7.02(t, J＝9.7Hz, 1H),6.58(dd, J＝9.1,3.8Hz, 1H),4.90(d, J＝6.4Hz, 1H),4.55(s, 1H),4.09(q, J＝5.2Hz, 1H),4.03(d, J＝8.8Hz, 1H),3.75(dt, J＝11.1,5.6Hz, 1H),3.68(d, J＝8.8Hz, 1H ),3.24(t,J＝10.9Hz,1H),3.17(d,J＝5.3Hz,2H),3.13–3.00(m,2H),2.88(dd,J＝7.5,4.4Hz,1H),2.82(m,J＝7.8,3.8Hz,1H),2.46(d,J＝7.6Hz,1H),1.93( m,J＝13.0,4.7Hz,1H),1.70(d,J＝13.5Hz,1H),0.69(m,J＝7.1,4.7Hz,2H),0.57–0.49(m,2H).

Embodiment 50

Compound D10 5-((3R, 4R)-3,4-dihydroxy-1-propionylpiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and propionyl chloride as raw materials, referring to the synthetic method of compound B4, compound D10 was obtained as a white solid with a yield of 30%. ¹ H NMR (400MHz, Chloroform-d) δ7.59 (s, 1H), 6.92 (t, J = 9.7 Hz, 1H), 6.49 (dd, J = 9.1, 3.9 Hz, 1H), 4.69–4.32 (m, 1H), 4.02–3.87 (m, 2H), 3.79 (d, J = 12.4 Hz, 1H), 3.67 (d, J = 13.5 Hz, 1H), 3.47–3 .19(m,1H),2.94(td,J＝7.6,3.0Hz,2H),2.88–2.69(m,2H),2.67–2.57(m,2H),2.47–2.31(m,2H),1.91(t,J＝12.4Hz,1H),1.73(ddd,J＝20.4,13.2,5 .3Hz,1H),1.16(td,J＝7.4,4.4Hz,3H).

Embodiment 51,

Compound D11 5-(((3R, 4R)-1-butyryl-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and butyryl chloride as raw materials, referring to the synthetic method of compound B4, compound D11 was obtained as a white solid with a yield of 25%. ¹ H NMR (400MHz, Chloroform-d) δ7.66 (s, 1H), 6.91 (td, J = 9.4, 2.4 Hz, 1H), 6.49 (dd, J = 9.1, 3.9 Hz, 1H), 4.65–4.34 (m, 1H), 4.02–3.86 (m, 2H), 3.80 (d, J = 12.6 Hz, 1H), 3.68 (d, J = 13.6 Hz, 1H), 3.51–3.37 (m, 1H), 3.26 (dd, J = 12.9,10.4Hz,1H),2.93(td,J＝7.5,4.1Hz,2H),2.81(dd,J＝10.2,5.9Hz,1H),2.61(q,J＝7.5Hz,2H),2.34(ddt,J＝10.0,7.1,3.9Hz,2H),1.90(t,J＝13.2Hz ,1H),1.66(dt,J＝14.7,6.0Hz,2H),0.97(td,J＝7.4,4.8Hz,3H).

Embodiment 52

Compound D12 5-(((3R, 4R)-1-(2-cyclopropylacetyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 2-cyclopropylacetyl chloride as raw materials, compound D12 was obtained as a white solid with a yield of 32% by referring to the synthetic method of compound B4. ¹ H NMR (400 MHz, Chloroform-d) δ7.61 (s, 1H), 6.97–6.85 (m, 1H), 6.49 (dd, J＝9.1, 3.9 Hz, 1H), 4.72–4.34 (m, 1H), 4.01–3.87 (m, 2H), 3.86–3.60 (m, 2H), 3.49–3.20 (m, 1H), 2.94 (td, J＝7.6, 3.4 Hz, 2 H),2.89–2.71(m,2H),2.62(q,J＝7.4Hz,2H),2.37–2.26(m,2H),1.91(dd,J＝16.4,4.7Hz,1H),1.75(td,J＝13.2,6.9Hz,1H),1.04(d,J＝7.3Hz,1H),0.57 (d, J＝7.9Hz, 2H), 0.19 (h, J＝4.7Hz, 2H).

Embodiment 53

Compound D13 5-(((3R, 4R)-1-(2-cyclohexylacetyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 2-cyclohexylacetyl chloride as raw materials, referring to the synthetic method of compound B4, compound D13 was obtained as a white solid with a yield of 22%. ¹ H NMR (400 MHz, Chloroform-d) δ7.62 (s, 1H), 6.92 (td, J = 9.5, 2.5 Hz, 1H), 6.48 (dd, J = 9.1, 3.9 Hz, 1H), 4.70–4.35 (m, 1H), 4.01–3.85 (m, 2H), 3.85–3.64 (m, 2H), 3.51–3.18 (m, 1H), 2 .93(q,J＝7.3Hz,2H),2.87–2.67(m,2H),2.62(q,J＝7.9Hz,2H),2.25(dd,J＝9.3,6.8Hz,2H),1.89(dd,J＝12.2,9.2Hz,1H),1.81–1.67(m,5H),1.34–1.09( m,5H),1.06–0.85(m,2H).

Embodiment 54

Compound D14 5-((3R, 4R)-1-(3-((difluoro-13-methyl)-12-fluorenyl)propanoyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 4,4,4-trifluorobutyryl chloride as raw materials, referring to the synthesis method of compound B4, compound D14 was obtained as a white solid with a yield of ¹³ %. NMR (400MHz, DMSO-d6) δ10.00(s,1H),7.01(t,J＝9.7Hz,1H),6.56(dd,J＝9.1,3.7Hz,1H),5.05(s,1H),4.71(s,1H),4.27(dd,J＝12.0,5.1Hz,1H),4.21–4.09( m,1H),4.06–3.97(m,1H),3.67(t,J＝8.5Hz,2H),3.62–3.46(m,2H),2.98–2.79(m,2H),2.75–2.53(m,2H),2.44(t,J＝7.5Hz,2H),2.38–2.25(m,2H), 1.80–1.58(m,2H).

Embodiment 55

Compound D15 5-(((3R, 4R)-3,4-dihydroxy-1-(propylsulfonyl)piperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and propylsulfonyl chloride as raw materials, referring to the synthetic method of compound B4, compound D15 was obtained as a white solid with a yield of 28%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.01(s, 1H),7.01(t, J＝9.7Hz, 1H),6.55(dd, J＝9.1,3.8Hz, 1H),5.11(d, J＝6.0Hz, 1H),4.70(s, 1H),4.02(d, J＝8.8Hz, 1H),3.67(dd, J＝14.0,7.2Hz, 2H),3.47–3.36(m, 2H ),3.04(m,J＝8.8,7.7,5.0Hz,3H),2.94–2.81(m,3H),2.46(d,J＝15.5Hz,2H),1.81(dd,J＝12.9,4.7Hz,1H),1.74–1.63(m,3H),1.00(t,J＝7.4Hz,2H).HR- MS:439.1315(M+Na) ⁺ .

Embodiment 56

Compound D16 5-(((3R, 4R)-1-(2-cyclopentylethyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 2-cyclopentylbromoethane as raw materials, referring to the synthetic method of compound B8, compound D16 was obtained as a white solid with a yield of 32%. ¹ H NMR (400MHz, Chloroform-d) δ7.58 (s, 1H), 6.91 (t, J = 9.4 Hz, 1H), 6.49 (dd, J = 9.1, 3.9 Hz, 1H), 4.03–3.86 (m, 3H), 2.98 (td, J = 7.7, 4.5 Hz, 2H), 2.82 (dd, J = 11.0, 4.4 Hz, 1H), 2.61(q,J＝6.6,5.5Hz,3H),2.46–2.36(m,2),2.33(t,J＝10.1Hz,1H),1.88–1.81(m,2H),1.74(dt,J＝14.3,8.9Hz,4),1.66–1.56(m,2),1.56–1.44(m, 4H),1.15–1.02(m,2H).

Embodiment 57

Compound D17 5-(((3R, 4R)-1-(2-cyclohexylethyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 2-cyclohexylbromoethane as raw materials, referring to the synthetic method of compound B8, compound D17 was obtained as a white solid with a yield of 37%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),7.00(dd,J＝10.4,9.0Hz,1H),6.54(dd,J＝9.1,3.8Hz,1H),4.60(d,J＝6.7Hz,1H),4.22(s,1H),3.94(d,J＝8.8Hz,1H),3.60(t,J＝7.9Hz,2H),2.86(m,J＝7.6,4.0Hz,2H),2.73-2.61(m,1H),2.46(t,J＝ 7.7Hz,2H),2.32(dd,J＝10.0,5.7Hz,2H),2.26–2.11(m,1H),2.08(t,J＝10.3Hz,1H),1.76(m,J＝13.1,4.4Hz,1H),1.72–1.52(m,6H),1.32(q,J＝7.3Hz,2 H),1.28–1.04(m,4H),0.89(q,J＝10.8,9.7Hz,2H).HR-MS:m/z 421.2498(M+H) ⁺ .

Embodiment 58

Compound D18 5-(((3R, 4R)-3,4-dihydroxy-1-(2-(tetrahydro-2H-pyran-4-yl)ethyl)piperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 4-(2-bromoethyl)tetrahydro-2H-pyran as raw materials, referring to the synthetic method of compound B8, compound D18 was obtained as a white solid with a yield of 26%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),7.00(t,J＝9.7Hz,1H),6.54(dd,J＝9.1,3.7Hz,1H),4.64(s,1H),4.27(s,1H),3.95(d,J＝8.7Hz,1H),3.86–3.74(m,2H),3.70–3.54(m,3H),3.26(m,J＝11.8,2.1Hz,2H),2.8 6(m,J＝7.6,4.0Hz,2H),2.74–2.55(m,1H),2.49–2.31(m,4H),2.18(d,J＝37.9Hz,1H),1.78(t,J＝12.9Hz,1H),1.63–1.32(m,6H),1.17(m,J＝16.2,6.7 ,4.5Hz,3H).HR-MS:m/z423.2296(M+H) ⁺ .

Embodiment 59

Compound D19 5-(((3R, 4R)-3,4-dihydroxy-1-(2-methoxyethyl)piperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 1-bromo-2-methoxyethane as raw materials and referring to the synthesis method of compound B8, compound D19 was obtained as a white solid with a yield of 10%. ¹ H NMR(400MHz,Chloroform-d)δ7.56(s,1H),6.91(t,J＝9.4Hz,1H),6.49(dd,J＝9.1,3.9Hz,1H),4.07–3.79(m,3H),3.53(t,J＝5.5Hz,2H),3.36(s,3H),2.98(t d,J＝7.7,4.3Hz,2H),2.90(dd,J＝10.6,4.3Hz,1H),2.74–2.58(m,5),2.50–2.41(m,1H),2.37(t,J＝10.2Hz,1H),1.86(dd,J＝6.8,4.4Hz,2H).

Embodiment 60

Compound D20 5-((3R, 4R)-3,4-dihydroxy-1-(2-(2-methoxyethoxy)ethyl)piperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 1-bromo-2-(2-methoxyethoxy)ethane as raw materials, referring to the synthesis method of compound B8, compound D20 was obtained as a white oil with a yield of ¹⁰ %. NMR(400MHz,Chloroform-d)δ7.59(s,1H),6.91(t,J＝9.4Hz,1H),6.49(dd,J＝9.1,3.9Hz,1H),4.02–3.89(m,3H),3.67–3.59(m,4H),3.58–3.53(m,2H),3.39(s, 3H),2.99(td,J＝7.6,3.5Hz,2H),2.90(dd,J＝11.1,4.3Hz,1H),2.70(t,J＝5.7Hz,2H),2.62(t,J＝7.7Hz,2H),2.56–2.49(m,1),2.44(t,J＝10.1Hz,1H),1 .87(q,J＝4.4Hz,3H).

Embodiment 61,

Compound D21 5-((3R, 4R)-1-(2-cyclopentylacetyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 2-cyclopentylacetyl chloride as raw materials, referring to the synthetic method of compound B4, compound D21 was obtained as a white solid with a yield of 48%. ¹ H NMR (400 MHz, Chloroform-d) δ7.59 (s, 1H), 6.90 (t, J = 9.5 Hz, 1H), 6.45 (dt, J = 9.0, 4.4 Hz, 1H), 5.80 (d, J = 25.9 Hz, 1H), 4.41 (s, 2H), 4.13 (s, 1H), 4.02 (d, J = 3.5 Hz, 1H), 3.76 (t, J = 5.7 Hz, 1H), 3.61 (t, J = 5.7 Hz,1H),3.06–2.93(m,2H),2.64(dd,J＝8.4,6.9Hz,2H),2.38(dd,J＝15.4,7.2Hz,2H),2.26(td,J＝17.3,16.3,8.6Hz,3H),1.86(q,J＝8.5,7.2Hz,2H),1.6 4–1.47(m,4H),1.17(td,J＝13.3,10.4,6.3Hz,2H).

Embodiment 62

Compound D22 5-((3R, 4R)-1-(butylsulfonyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and butylsulfonyl chloride as raw materials, referring to the synthetic method of compound B4, compound D22 was obtained as a white solid with a yield of 62%. ¹ H NMR (400 MHz, Chloroform-d) δ7.60 (s, 1H), 6.92 (t, J = 9.4 Hz, 1H), 6.49 (dd, J = 9.1, 3.9 Hz, 1H), 4.03-3.90 (m, 3H), 3.78 (ddd, J = 11.7, 5.4, 1.9 Hz, 1H), 3.68-3.60 (m, 1H), 3.16 ( ddd,J＝12.2,10.3,4.8Hz,1H),3.03–2.89(m,5H),2.72(d,J＝5.9Hz,1H),2.68–2.58(m,3H),1.98–1.90(m,2H),1.87–1.76(m,2H),1.47(h,J＝7.4Hz,2 H),0.97(t,J=7.4Hz,3H).

Embodiment 63

Compound D23 5-((3R, 4R)-1-((3-chloropropyl)sulfonyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 3-chloropropanesulfonyl chloride as raw materials, referring to the synthetic method of compound B4, compound D23 was obtained as a white solid with a yield of 44%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),7.01(dd,J＝10.4,9.0Hz,1H),6.54(dd,J＝9.1,3.8Hz,1H),5.12(d,J＝6.0Hz,1H),4.73(d,J＝1.0Hz,1H),4.01(d,J＝8.8Hz,1H),3.75(t,J＝6.5Hz,2H),3.70–3.61(m,2H) ,3.43(dd,J＝11.1,5.7Hz,2H),3.23–3.15(m,2H),3.12–3.00(m,1H),2.95–2.81(m,3H),2.45(t,J＝7.6Hz,2H),2.18–2.04(m,2H),1.83(td,J＝13.3, 4.8Hz,1H),1.71(d,J＝13.9Hz,1H).

Embodiment 64

Compound D24 5-((3R, 4R)-1-(sec-butylsulfonyl)-3,4-dihydroxypiperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and sec-butylsulfonyl chloride as raw materials, referring to the synthetic method of compound B4, compound D24 was obtained as a white solid with a yield of 49%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.00(s,1H),7.01(dd,J＝10.4,9.1Hz,1H),6.55(dd,J＝9.1,3.8Hz,1H),5.08(dd,J＝6.0,1.1Hz,1H),4.68(d,J＝1.1Hz,1H),4.00(d,J＝8.8Hz,1H),3.62(dd,J＝14.3,6.9Hz,2H),3.47(dt,J＝13.0,6.8Hz,2H),3.18–3 .05(m,2H),2.96(td,J＝11.2,5.2Hz,1H),2.86(td,J＝7.6,3.9Hz,2H),2.45(t,J＝7.7Hz,2H),1.92–1.72(m,2H),1.67(d,J＝13.7Hz,1H),1.42(ddd,J＝13 .8,9.2,7.3Hz,1H),1.20(d,J＝6.8Hz,3H),0.95(t,J＝7.5Hz,3H).

Embodiment 65

Compound D25 5-((3R, 4R)-3,4-dihydroxy-1-((3,3,3-trifluoropropyl)sulfonyl)piperidin-4-yl)methoxy)-8-fluoro-3,4-dihydroquinolin-2(1H)-one

Using intermediate 20 and 3,3,3-trifluoropropane-1-sulfonyl chloride as raw materials, referring to the synthetic method of compound B4, compound D25 was obtained as a white solid with a yield of 66%. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.01(s, 1H),7.01(dd,J＝10.5,9.1Hz,1H),6.55(dd,J＝9.1,3.7Hz,1H),5.12(d,J＝6.0Hz,1H),4.72(d,J＝1.0Hz,1H),4.01(d,J＝8.8Hz,1H),3.72–3.60(m,2H),3.45(dd,J＝11.0,5.4Hz,2H) ,3.35(dd,J＝6.9,4.2Hz,2H),3.16–3.07(m,1H),2.97(t,J＝11.0Hz,1H),2.86(td,J＝7.6,4.0Hz,2H),2.75–2.63(m,2H),2.45(t,J＝7.6Hz,2H),1.83(t d,J＝13.1,4.7Hz,1H),1.74–1.65(m,1H).

The beneficial effects of the present invention are demonstrated by experimental examples below.

Experimental Example 1: In vitro anti-mycobacterium tuberculosis activity test of compounds

1. In vitro anti-Mycobacterium tuberculosis H37Ra activity test of compounds

Add H37Ra (ATCC 25177) strain to 7H9+ADN medium, culture on a 160rpm shaker for 2 weeks until the logarithmic growth phase, dilute and adjust the bacterial solution to about 1*10 ⁵ CFU/mL for later use. Dissolve the test compound in DMSO to prepare a high concentration solution of 10 mg/mL, and store at 2-8°C. Take samples and dilute the above bacterial solution to 10-50 μg/mL, add two replicates of each compound, and add 8-10 concentration gradients (2-fold gradient dilution) to a 96-well plate in sequence. Use an equal amount of blank DMSO solution as a blank control and a bacterial solution without drug addition as a growth control. The volume of each well is 200 μL. After adding the plate, 5% CO ₂ was placed at 37°C for 5 days to check the growth of tuberculosis bacilli. 1% Resazurin solution was added to the growth control wells for overnight culture to observe whether the color has changed from blue to red. At this time, 1% Resazurin solution was added to all the remaining wells for overnight culture, and the fluorescence value at 540/590nm was measured using a PE Envision multifunctional microplate analyzer. The lowest drug concentration corresponding to a mortality rate of more than 90% of the strain measured by fluorescence absorption is the MIC value of the compound for H37Ra, and the calculation formula is: mortality rate = 1-(fluorescence value of the test well-fluorescence value of the blank control)/(fluorescence value of the growth control-fluorescence value of the blank control).

The inhibitory activity of the compounds against Mycobacterium tuberculosis H37Ra is shown in Table 1:

Table 1 Results of the anti-Mycobacterium tuberculosis H37Ra activity of the compounds

Note: A: MIC<0.001μM; B: 0.001μM≤MIC<0.01μM; C: 0.01μM≤MIC<0.1μM; D: 0.1μM≤MIC<1μM; E: MIC≥1μM.

The results in Table 1 show that most compounds have significant inhibitory activity against Mycobacterium tuberculosis H37Ra in vitro. Among them, the antibacterial activities of C3, C4, D2 and D17 are comparable to those of the positive control OPC-167832, and the MIC values are lower than 0.001 μM, showing significant antibacterial activity.

2. In vitro anti-Mycobacterium tuberculosis H37Rv activity test of compounds

Compound solution preparation: The compounds to be tested will be tested starting at 64 μg/mL. Dissolve the compounds in DMSO at a concentration of 2048 μg/mL as a frozen stock solution (-80°C). Add 100 μL of the stock solution to 700 μL 7H9 w/o Tween for an 8-fold dilution to make 800 μL of a 256 μg/mL solution for setting up the well plate.

96-well plate setup: The culture medium is 7H9, OD600=0.001 type culture medium, 100 μL of test compound or positive control is added to two columns in duplicate (for duplicate, add four times). Take 100 μL from the second column to the third column and mix well, which is a two-fold serial dilution, and mix it thoroughly by blowing up and down 8 times. Continuously dilute to the 11th column and discard the last 100 μL of solution. After adding the drug, seal and culture at 37°C for 7 days. Detect the drug concentration corresponding to the death of more than 90% of the strains and record the MIC value.

The inhibitory activity of the main compounds against Mycobacterium tuberculosis H37Rv is shown in Table 2:

Table 2 Anti-Mycobacterium tuberculosis H37Rv activity results of compounds

Note: A: MIC<0.1μM; B: 0.1μM≤MIC<1μM; C: MIC≥1μM.

The results in Table 2 show that for the compounds with better anti-Mycobacterium tuberculosis H37Ra activity, the in vitro anti-Mycobacterium tuberculosis H37Rv activity was consistent with that of the positive compounds, and both exhibited excellent anti-tuberculosis activity.

3. In vitro activity test of compounds against drug-resistant Mycobacterium tuberculosis (MDR-TB and XDR-TB) Dosing:

Test compound: completely dissolved in DMSO, prepared into a storage stock solution, filtered and sterilized, then serially diluted with Middlebrook 7H9 culture medium of Mycobacterium tuberculosis, added to 96-well culture plate at 100 μL/well before use, the final drug concentration is: 1 μg/mL, 0.5 μg/mL, 0.25 μg/mL, 0.125 μg/mL, 0.0625 μg/mL, 0.03125 μg/mL, 0.0156 μg/mL, 0.0078 μg/mL, 0.0039 μg/mL, 0.00019 μg/mL, 0.0001 μg/mL.

Control: Add DMSO containing the same concentration as the highest concentration of the test drug.

Tuberculosis: MDR/XDR strains (Y117, Y16)

Take the strain that has been passaged 1-2 times and is in the logarithmic growth phase, grind the bacteria to OD600 = 1.0 (about 5×10 ⁶ CFU/mL), take 200 μL and dilute it into 10 mL of culture medium, then add it to the culture plate with a continuous pipette at 100 μL/well (about 10 ⁴ CFU/well); add the same amount of bacteria as the test compound (D100), 1/10 of the amount of bacteria (D10) and 1/100 of the amount of bacteria (D1) to the control group.

Observation results:

After culturing at 37°C for 14 days, the growth of colonies in each group was observed, and the lowest concentration of the drug group with no colony growth was taken as the MIC value of the test compound for the strain.

The inhibitory activities of the main compounds against drug-resistant Mycobacterium tuberculosis MDR-TB (Y117) and XDR-TB (Y16) are shown in Table 3:

Table 3 Activity results of compounds against drug-resistant Mycobacterium tuberculosis Y117 and Y16

Note: A: MIC ₁₀₀ <0.01μg/mL; B: 0.01μg/mL≤MIC ₁₀₀ <0.05μg/mL; C: 0.05μg/mL≤MIC ₁₀₀ <0.1μg/mL; D: MIC ₁₀₀ ≥0.1μg/ mL

The above results indicate that compounds C4 and D17 exhibit significant bactericidal activity against clinically isolated Mycobacterium tuberculosis and have good clinical application value.

Experimental Example 2: Compound Solubility Test

Experimental Procedure

1. Prepare pure water

2. Sample preparation: Add 8 μL of 10 mM control solution and test compound stock solution to 792 μL of water.

3. Shake the sample tube at room temperature for 1 h (1000 rpm).

4. Preparation of standard curve

4.1 Preparation of 300 μM spiking solution (SS): Add 6 μL of 10 mM compound to 194 μL of MeOH/ACN (4/1) solution.

4.2 Prepare a standard curve in MeOH/ACN (4/1) solution.

5. Centrifuge the sample (10 min - 12000 rpm) to precipitate undissolved particles and transfer the supernatant to a new sample bottle.

6. Dilute the supernatant 10 times and 100 times with water

Add 10 μL of supernatant to 90 μL of water to make a 10-fold dilution.

10 μL of supernatant was added to 990 μL of water to make a 100-fold dilution.

7. Sample Preparation for LC-MS/MS

5 μL of samples (no dilution, 10-fold dilution, and 100-fold dilution) and standard curve samples were added to 95 μL of CAN and loaded for analysis.

The solubility data of the main compounds are shown in Table 4.

Table 4 Solubility of compounds

The above results show that compared with the positive control, the solubility of compounds D17 and D24 is significantly improved, indicating that the compounds may have better pharmacokinetic properties and potential clinical application value.

In summary, the present invention provides a new quinolinone derivative with excellent anti-Mycobacterium tuberculosis activity. As a new type of DprE1 enzyme inhibitor, it has controllable toxicity, excellent anti-tuberculosis activity and pharmacokinetic properties, and has very good clinical application prospects.

Claims (13)

1. A quinolinone derivative represented by formula I or a pharmaceutically acceptable salt or solvate thereof:

Wherein, Ring A is an aryl group, an aromatic heterocyclic group or a paraaromatic heterocyclic group; L is O or NR ₇ ; L' and L" are independently selected from none, CO, SO ₂ , NH or (CH ₂ ) _m , m is an integer of 1 to 3; M ₁ and M ₂ are independently selected from N or CH; R ₁ , R ₂ , and R ₃ are independently selected from H, halogen, C1-C3 alkoxy, hydroxyl, -NR ₁₁ R ₁₂ , or R ₂ and R ₃ are connected to form a 3-7 membered ring; R ₁₁ is H or C1-C3 alkyl, R ₁₂ is C1-C3 alkyl or -CO-R ₁₃ , and R ₁₃ is halogen substituted or unsubstituted C1-C3 alkyl; _R4 is -H or -OH; R ₅ and R ₆ are independently selected from -H, -Boc _, a bridged hydrocarbon group, Ra, _Rb , _Rc substituted or unsubstituted aryl, halogen substituted or unsubstituted aromatic heterocyclic group, _Rd substituted or unsubstituted C1-C4 alkyl, C3-C6 cycloalkyl or

t is an integer from 1 to 3; wherein _Ra , _Rb , and _Rc are independently selected from halogen, nitro,

Wherein, R', R" are independently selected from: H, -CO- _Rs or _{-SO2- Rs} ; R"' is _-ORs or

R _s , R _p , and R _q are independently selected from H, C1-C3 alkyl, or R _p and R _q are connected to form a ring; R _d is halogen, halogen-substituted or unsubstituted C1-C3 alkyl, C3-C6 cycloalkyl or C3-C6 heterocycloalkyl; _R7 is a C1-C3 alkyl group. 2. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 1, wherein ring A is

3. The quinolinone derivative according to claim 1 or 2, or a pharmaceutically acceptable salt or solvate thereof, characterized in that _R1 , _R2 , and _R3 are independently selected from H, F, methoxy, hydroxyl, _-NR11R12 , or _R1 and _R2 are connected to form a three-membered ring; _R11 is H or methyl, _R12 is methyl or -CO- _R13 , _{and R13} is a methyl group substituted or unsubstituted with 1 to 3 Fs. 4. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 3, characterized in that _R1 is H, F or methoxy, _R2 is _H or _-NR11R12 , _R3 is H, or _R2 and _R3 are connected to form a saturated 3-membered carbocyclic ring; _R11 is H or methyl, _R12 is methyl or -CO- _R13 , and _R13 is methyl or _-CF3 . 5. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 4, characterized in that the quinolinone derivative has a structure shown in Formula II-A:

Preferably, R ₁ is H or F, R ₂ is H and R ₃ is H, or R ₂ and R ₃ are connected to form a saturated 3-membered carbon ring. 6. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 4, wherein the quinolinone derivative has a structure as shown in Formula II-B:

wherein R ₃ is H; Preferably, R ₁ is H or F, R ₂ is -NR ₁₁ R ₁₂ ; R ₁₁ is H or methyl, R ₁₂ is methyl or -CO-R ₁₃ , and R ₁₃ is methyl or -CF ₃ . 7. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 4, wherein the quinolinone derivative has a structure shown in Formula II-C or Formula II-D:

Wherein, R ₂ and R ₃ are both H; Preferably, R ₁ is H, F or methoxy. 8. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 4, wherein the quinolinone derivative has a structure shown in Formula II-E or Formula II-F:

Wherein, R ₁ , R ₂ , and R ₃ are all H. 9. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to any one of claims 1 to 8, characterized in that the quinolinone derivative has a structure shown in Formula III-A, Formula III-B, Formula III-C or Formula III-D:

10. The quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to claim 9, wherein the quinolinone derivative has the following structure:

11. A method for synthesizing the quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to any one of claims 1 to 10, characterized in that it comprises the following steps: reacting compound A and compound B at 70 to 90° C. for 4 to 8 hours under the action of an inorganic base to obtain a compound of formula I; The reaction formula is as follows:

Wherein, R ₀ is -OH or -NH-R ₇ . 12. Use of the quinolinone derivative according to any one of claims 1 to 11 or a pharmaceutically acceptable salt or solvate thereof as a DprE1 enzyme inhibitor. 13. Use of the quinolinone derivative or a pharmaceutically acceptable salt or solvate thereof according to any one of claims 1 to 11 in the preparation of a drug for treating tuberculosis; preferably, the drug is an anti-Mycobacterium tuberculosis drug; more preferably, the drug for treating tuberculosis is a drug for treating drug-resistant tuberculosis, and the Mycobacterium tuberculosis is drug-resistant Mycobacterium tuberculosis.