CN108047145B - 2-arylquinoxaline compound with affinity with Tau protein and preparation method and application thereof - Google Patents

Abstract

The invention provides a 2-arylquinoxaline compound with affinity with Tau protein, which has a structure shown in a formula (I):

Description

2-arylquinoxaline compound with affinity with Tau protein and preparation method and application thereof

Technical Field

The invention relates to the technical field of radiopharmaceutical chemistry and clinical nuclear medicine, in particular to a 2-arylquinoxaline compound with high affinity with Tau protein and a preparation method and application thereof.

Background

The progressively developing fatal neurodegenerative Disease Alzheimer's Disease (AD) has become the fourth largest Disease that seriously threatens the health of the elderly, following tumors, heart Disease and stroke and diabetes. Its clinical manifestations are mainly cognitive, memory and life capacity decline, accompanied by neuropsychiatric symptoms and behavioral disorders. Lancet reported in 2013, 193 million AD patients in 1990, 371 million in 2000, and 596 million in 2010. It is predicted that the population of AD patients worldwide will range from 2610 ten thousand in 2006 to 1.1 million in 2050. Therefore, the method has extremely important significance for the prevention of AD and the research of early diagnosis.

It is a complex of factors such as age, genetics, environment, brain trauma, depression, diabetes, hyperlipidemia, vascular disease, etc. that contribute to AD. Excluding the complexity of AD itself, its early accurate diagnosis and the lack of detection means also hampers its effective treatment. At present, the clinical diagnosis of AD can only depend on the autopsy of the patient after death. Autopsy results reveal two major pathological features of AD: 1) deposition of Senile Plaques (SPs) composed of Α β polypeptides; 2) neurofibrillary tangles (NFTs) composed of highly phosphorylated Tau protein. Therefore, the method has great application value for quantification of the A beta polypeptide in the brain and the protein level of Tau. Modern molecular imaging has the advantages of non-invasive, accurate diagnosis and monitoring of disease progression, and has gained tremendous progress in recent years. Positron Emission Tomography (PET) scanning imaging in particular is a very powerful tool in drug development, monitoring the course of disease changes. PET imaging agents targeting ap deposition have shown a great deal of clinical data, however, ap deposition density in brain tissue has no good correlation with neurodegeneration and dysesthesia, and may result in false positive imaging results. The Tau protein density in the brain and spread in the cerebral cortex are well correlated with a β deposition. Therefore, Tau protein is an ideal target for AD diagnosis. Other studies have also shown that Tau protein is a central factor in the pathogenesis of AD.

There are many international reports on the use of Tau protein as a target point of PET probes. Of the THK series, THK-5105(Okamura, N et al. journal of Nuclear medicine.2013,54: 1420-. However, it has high nonspecific binding in brainstem, thalamus and subcortical white matter, and the slow clearance rate in vivo further limits its clinical application. Again modified THK-5117, THK-5331 is currently in clinical research only. PBB3(Maruyama, M et al. neuron.2013,79:1-15) has high affinity for Tau protein and high selectivity relative to a β protein, but it has in vivo instability due to its own chemical structure with cis-trans configuration. T808(Cashion, D et al. International Patent WO 2011/119565A1,2011), which satisfies almost all the requirements of a Tau probe, has severe defluorination in vivo, preventing further clinical development, and its analogue T807 is currently in the experimental stage.

Disclosure of Invention

The invention aims to provide a 2-arylquinoxaline compound with high affinity with Tau protein, and a preparation method and application thereof.

In order to achieve the purpose of the invention, the invention provides a 2-aryl quinoxaline compound with affinity with Tau protein, and the structure of the compound is shown as the formula (I):

wherein X and Y each independently represent N or CH; r₁At position 4-7, R₂At the 2 ' position, the 3 ' position or the 4' position.

R₁And R₂Each independently represents H,¹⁸F、¹⁹F、¹²³I、¹²⁵I、¹²⁷I、OH、O¹¹CH₃、O¹²CH₃、NH₂、NH¹¹CH₃、NH¹²CH₃、N(CH₃)₂、

O(CH₂)_m ¹⁸F or O (CH)₂)_m ¹⁹F. Wherein R is OH,¹⁸F or¹⁹F and m are integers between 1 and 6.

Preferably, the compound of the present invention is any one of the following formulas 1) to 13):

wherein F in the formulae 4) to 13) is¹⁸F or¹⁹F。

The preparation method of the compound of the invention comprises the following steps:

with X and Y being CH, R₁Is 5 positionOr 6-position N (CH)₃)₂，R₂For the case of the 4' -OH, i.e., the compounds of formula 1) and 2), other different substituents or different substitution positions are prepared in a similar manner:

(1) take 10mmol

Dissolved in 10mL of DMSO, stirred at room temperature for 30 minutes, and then 10mmol of DMSO was slowly added thereto under ice-bath conditions

After the reaction was continued for 2 hours after the temperature of the DMSO solution was raised to room temperature, 200mL of saturated NaHCO was added₃Precipitating yellow precipitate in the solution, performing suction filtration, washing with a large amount of water, and drying to obtain a 5-position or 6-position substituted mixture;

(2) dissolving 1.0g of the mixture and 2.0g of oxalic acid dihydrate in 200mL of absolute ethanol under a heating condition to obtain a mauve solution, cooling to separate out a solid, performing suction filtration to obtain 6-substituted mauve oxalate crystals, dropwise adding petroleum ether into the filtrate to separate out a solid, performing suction filtration to obtain 5-substituted dark red oxalate crystals, respectively adding 5.0mL of strong ammonia water into the crystals to obtain yellow solids, performing suction filtration, washing and drying to obtain the compounds shown in the formulas 1) and 2).

With X and Y being CH, R₁Is a 6-substituted N (CH)₃)₂，R₂Is O (CH) at the 4' position₂)₂ ¹⁸F、

For example, compounds described by formula 3), formula 6) or formula 11), other different carbon chain lengths, different optical configurations, different substituents or different substitution positions are prepared in a similar manner:

(1) taking 1mmol of the compound shown in the formula 1), 1mmol of CsF and 1.2 mmol of

Dissolving in 5mL of anhydrous DMF, stirring at 90 deg.C for 2 hr, adding 50mL of ice water to precipitate out a large amount of yellow precipitate, and pumpingFiltering, washing and drying to respectively obtain compounds shown as a formula (V), a formula (VI) or a formula (VII);

(2) dissolving 1mmol of a compound shown as a formula (VI) in 20mL of THF, adding 10mL of 1M HCl, heating at 60 ℃ for reaction for 1 hour, then performing rotary evaporation to remove the THF, adding 5.0mL of ammonia water to precipitate yellow, performing suction filtration, washing and drying to obtain a compound shown as a formula (VIII);

(3) dissolving 1mmol of a compound shown as a formula (VIII) in 10mL of anhydrous dichloromethane, adding 1mmol of p-toluenesulfonyl chloride and 10mmol of triethylamine, reacting at room temperature for 12 hours, removing the solvent by rotary evaporation, and performing column chromatography separation to obtain a compound (yellow solid) shown as a formula (IX);

(4) dissolving 1mmol of the compound shown in the formula (IX) in 10mL of anhydrous dichloromethane, adding 4mmol of 3, 4-dihydropyran and 1.0g of pyridine p-toluenesulfonate (PPTS), stirring at 50 ℃ for reaction for 12 hours, removing the solvent by rotary evaporation after the reaction is finished, and separating by column chromatography to obtain the compound shown in the formula (X) (oily liquid);

(5) 1-5mg of the compound (V) or (VII) or (X) is dissolved in 1mL of anhydrous acetonitrile, which is added to the K-containing solution which has been freed of water₂₂₂/K₂CO₃5-30mCi¹⁸F^-In the reaction tube (2), labeling is carried out for 5min at the temperature of 100 ℃; for the compounds (VII) and (X), 100. mu.L of 1M HCl is added, and after further reaction at 100 ℃ for 5min, saturated NaHCO is added separately₃Neutralizing the solution; finally, the mixture is passed through a C18 reverse phase column (Sep-pak), washed with water to remove salts and the remainder¹⁸F^-Then respectively leaching with acetonitrile to obtain final products, and processing by N₂After blowing dry, the mixture was separated by HPLCRespectively obtaining the compound shown in the formula (V), the formula (VI) or the formula (VII) with the purity of more than 98 percent.

The derivatives of the compounds shown in the formula (I), such as pharmaceutically acceptable salts, esters or amides or prodrugs, all belong to the protection scope of the invention.

The present invention also provides a diagnostic or detection reagent for a neurofibrillary tangle disease caused by Tau protein deposition, the active ingredient of which is a compound represented by the formula (I), and/or a derivative thereof.

These diseases include, but are not limited to, alzheimer's disease, frontotemporal lobar degeneration, chronic traumatic encephalopathy, progressive supranuclear palsy, corticobasal degeneration, or pick's disease, among others.

The invention further provides application of the compound shown in the formula (I) and/or derivatives thereof in preparing nuclear medicine imaging agents (such as bifunctional imaging agents) or optical imaging agents.

Compared with the prior art, the invention has the following advantages:

the compound of formula (I) provided by the invention can be directly used as a fluorescent probe for detecting neurofibrillary tangles in vivo or in a tissue sample by using 1 mu M¹⁹The staining time is fast and efficient when staining tissue sections with the solution of compound F (neurofibrillary tangles in the sections can be stained efficiently in 30 seconds and do not require washing) and when used for nuclear medicine imaging, they are labelled with a suitable radioisotope. The compounds are particularly useful for diagnosing patients with neurofibrillary tangles (Tau protein deposits) characteristic of Alzheimer's disease.

Drawings

FIG. 1 is a schematic diagram of the synthesis of the compounds of examples 1-21 of the present invention; wherein the reaction reagents and conditions involved are: (a) dimethyl sulfoxide, room temperature, 10 hours; (b) hydrazine hydrate, palladium carbon and methanol are refluxed for 2 hours at 90 ℃; (c) paraformaldehyde, sodium methoxide, sodium borohydride and methanol are refluxed for 5 hours at 90 ℃; (d) (R) -glycidyl-3-nitrobenzenesulfonate, cesium fluoride, nitrogen dimethylformamide, 65 ℃,6 hours; (e) refluxing tetrabutylammonium fluoride and anhydrous tetrahydrofuran for 6 hours; (f) (R) - (-) -p-methylbenzenesulfonic acid-2, 2-dimethyl-1, 3-dioxolaneCyclyl-4-methyl ester, cesium fluoride, azodicarbonamide, at 65 ℃ for 6 hours; (g) hydrochloric acid (1M), refluxing for 30 min at 90 ℃, sodium bicarbonate; (h) tert-butyldimethylsilyl chloride, imidazole, acetonitrile, room temperature, 12 hours; (i) di-tert-butyl dicarbonate and anhydrous tetrahydrofuran are refluxed for 48 hours at 90 ℃; (j) refluxing tetrabutylammonium fluoride and anhydrous tetrahydrofuran for 3 hours; (k) p-toluenesulfonyl chloride, triethylamine and dichloromethane at room temperature for 6 hours; (l)3, 4-dihydro-2H-pyran, 4-methylbenzenesulfonic acid pyridine (PPTS), dichloromethane, room temperature, 5 hours; (m) (1)¹⁸F-ion, 4,7,13,21, 24-hexaoxa-1, 10-diazabicyclo [8,8]-hexacosane, potassium carbonate, at 100 ℃, for 6 minutes, (2)1M hydrochloric acid, at 100 ℃, for 5 minutes; (n) (S) -glycidyl-3-nitrobenzenesulfonate, cesium fluoride, nitrogen dimethylformamide, 65 ℃,6 hours; (o) (S) - (-) -p-methylbenzenesulfonic acid-2, 2-dimethyl-1, 3-dioxolanyl-4-methyl ester, cesium fluoride, azodicarbonamide, at 65 ℃ for 6 hours.

FIG. 2 is a schematic diagram of the synthesis of the compounds of examples 22-48 of the present invention; wherein the reaction reagents and conditions involved are: (a) nitrogen-nitrogen dimethylformamide, potassium carbonate, 150 ℃, and reflux for 12 h; (b) hydrogen, palladium carbon and methanol are carried out for 6 hours at room temperature; (2) dimethyl sulfoxide, room temperature, 10 hours; (c) (R) -glycidyl-3-nitrobenzenesulfonate, cesium fluoride, nitrogen dimethylformamide, 65 ℃,6 hours; (d) refluxing tetrabutylammonium fluoride and anhydrous tetrahydrofuran for 6 hours; (e) (R) - (-) -p-methylbenzenesulfonic acid-2, 2-dimethyl-1, 3-dioxolanyl-4-methyl ester, cesium fluoride, azodicarbonamide, at 65 ℃, for 6 hours; (f) hydrochloric acid (1M), refluxing for 30 min at 90 ℃, sodium bicarbonate; (g) p-toluenesulfonyl chloride, triethylamine and dichloromethane at room temperature for 6 hours; (h)3, 4-dihydro-2H-pyran, 4-methylbenzenesulfonic acid pyridine (PPTS), dichloromethane, room temperature, 5 hours; (i) (1)¹⁸F-ion, 4,7,13,21, 24-hexaoxa-1, 10-diazabicyclo [8,8]-hexacosane, potassium carbonate, at 100 ℃, for 6 minutes, (2)1M hydrochloric acid, at 100 ℃, for 5 minutes; (j) (S) -glycidyl-3-nitrobenzenesulfonate, cesium fluoride, nitrogen dimethylformamide, 65 ℃,6 hours; (k) (S) - (-) -p-methylbenzenesulfonic acid-2, 2-dimethyl-1, 3-dioxolanyl-4-methyl ester, cesium fluoride, azodicarbonamide, at 65 ℃ for 6 hours.

FIG. 3 is a schematic representation of the synthesis of the compounds of examples 49-63 of the present inventionAn intent; wherein the reaction reagents and conditions involved are: (a) (4R,5R) -4, 5-bis (tosyloxymethyl) -2, 2-dimethyl-1, 3-dioxolane, cesium fluoride, azodicarbonamide, at 65 ℃ for 6 hours; (b) refluxing tetrabutylammonium fluoride and anhydrous tetrahydrofuran for 6 hours; (c) hydrochloric acid (1M), refluxing for 30 min at 90 ℃, sodium bicarbonate; (d) (4S,5S) -4, 5-bis (tosyloxymethyl) -2, 2-dimethyl-1, 3-dioxolane, cesium fluoride, azodicarbonamide, at 65 ℃ for 6 hours; (e) (1)¹⁸F-ion, 4,7,13,21, 24-hexaoxa-1, 10-diazabicyclo [8,8]-hexacosane, potassium carbonate, 100 ℃,6 minutes, (2)1M hydrochloric acid, 100 ℃,5 minutes.

FIG. 4 is a schematic diagram of the synthesis of compounds of examples 64-87 of the present invention; wherein the reaction reagents and conditions involved are: (a) dimethyl sulfoxide, room temperature, 10 hours; (b) 1-bromo-2-fluoroethane, potassium carbonate, nitrogen dimethylformamide, 90 ℃; (c) 10% palladium on carbon, hydrazine hydrate, methanol, 80 ℃; (d) methyl iodide, acetone, 40 deg.C, water.

FIG. 5 shows the results of fluorescent staining of brain tissue sections of AD patients with S-5, R-5, S-16, R-16, S-23 and R-23 probes, respectively, according to example 89 of the present invention; GFP channel is the staining of the compound of the invention and RFP channel is the staining of the a β plaque specific probe.

FIG. 6 is S-, [ 2 ] in example 90 of the present invention¹⁸F]23 autoradiography of brain tissue sections from AD patients (A, C and E), and results were confirmed by thioflavin-S staining (B, D and F). C and D are enlarged views.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.

Example 1: synthesis of intermediate 1

The compound 4-nitrophthaldiamine (3.1g,20.0mmol) was dissolved in a 250mL round bottom flask with 8mL DMSO, then 2-bromo-4' -hydroxyacetophenone (4.3g,20.0mmol) was slowly added to the reaction flask, stirred at room temperature for 10 hours, after the reaction was complete, a yellow solid precipitated, the precipitated product was filtered with suction and washed with waterWashing with 100mL of deionized water and 50mL of methanol, and drying the solid obtained by suction filtration to obtain 3.1g of intermediate 1, wherein the yield is 58.0%, and the structure is as follows: MS m/z calcd for C₁₄H₁₀N₃O₃ 268.1；found,M+H+.

Example 2: synthesis of intermediate 2

Dissolving intermediate 1(4.3g,16.1mmol) and palladium carbon (0.8g,16.1mmol) in 150mL of methanol in a 250mL round-bottom flask, slowly adding hydrazine hydrate (5.1g,48.0mmol) dropwise to the reaction solution, heating and refluxing for 1 hour by using a 90 ℃ oil bath, after the reaction is finished, removing the palladium carbon by suction filtration while hot, and distilling under reduced pressure to remove the methanol to obtain a crude product, and recrystallizing the crude product with methanol to obtain 3.4g of white intermediate 2, wherein the yield is 89.0%, and the structure is as follows: MS m/z calcd for C₁₄H₁₂N₃O 238.1；found,M+H+.

Example 3: synthesis of intermediate 3

Dissolving the intermediate 2(1.2g,5.0mmol) with 50mL of methanol, sequentially adding paraformaldehyde (0.6g,20.0mmol) and sodium methoxide (2.0mL,10.0mmol), heating and refluxing in a 90 ℃ oil bath for 2 hours, slowly adding sodium borohydride (4.3g,16.1mmol) after the reaction liquid is cooled, continuing heating and refluxing at 90 ℃ for 3 hours, adjusting the pH of the reaction liquid to be neutral by using 1M HCl after the reaction is finished, removing the methanol by reduced pressure distillation, adding 100mL of deionized water into a round bottom flask, performing ultrasonic treatment for half an hour, performing suction filtration, drying the obtained compound to obtain 1.2g of intermediate 3, wherein the yield is 95.5%, and the structure is as follows:¹H NMR(400MHz,DMSO-d₆)9.78(s,1H),9.16(s,1H),8.03(d,J＝8.2Hz,2H),7.71(d,J＝9.1Hz,1H),7.23(d,J＝9.1Hz,1H),6.89(d,J＝8.3Hz,2H),6.71(s,1H),6.59(d,J＝4.4Hz,1H),2.81(d,J＝4.4Hz,3H).MS:m/z calcd for C₁₅H₁₄N₃O 252.1；found 252.2,M+H+.

example 4: synthesis of intermediate (R) -4

Intermediate 3(0.5g,2.0mmol) and (R) -glycidyl-3-nitrobenzenesulfonate (0.5g,2.5mmol) were dissolved in 5mL of DMF and cesium fluoride (0.9g,6.0mmol) was added with stirring and reacted at 65 ℃ for 6 hours. After the reaction is finished, after the reaction solution is cooled to room temperature, 100mL of deionized water with yellow color is addedSeparating out the color product, placing the color product in ultrasonic to act for 30 minutes, and performing suction filtration to obtain a yellow crude product. After drying the crude product, column chromatography separation is carried out by using a developing solvent with the volume ratio of petroleum ether to ethyl acetate being 2:1, so as to obtain 365.1mg of light yellow solid. Yield 59.4%, structure is as follows:¹H NMR(400MHz,CDCl₃)9.05(s,1H),8.13–7.98(m,2H),7.89(d,J＝9.1Hz,1H),7.17(dd,J＝9.1,2.4Hz,1H),7.12–7.01(m,2H),4.32(dd,J＝11.0,3.1Hz,1H),4.04(dd,J＝11.0,5.7Hz,1H),3.40(ddd,J＝7.0,5.7,2.9Hz,1H),3.02(s,3H),2.98–2.85(m,1H),2.80(dd,J＝4.9,2.6Hz,1H).MS:m/z calcd for C₁₈H₁₈N₃O₂ 308.1；found 308.3,M+H+.

example 5: synthesis of intermediate (S) -4

Intermediate (S) -4 was obtained from intermediate 3 according to the procedure for the synthesis of intermediate (R) -4 to give 378.4mg as a pale yellow solid. Yield 61.6%, structure as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.14–7.99(m,2H),7.88(d,J＝9.1Hz,1H),7.15(dd,J＝9.1,2.5Hz,1H),7.07(d,J＝8.9Hz,2H),7.01(d,J＝2.2Hz,1H),4.31(dd,J＝11.0,3.1Hz,1H),4.04(dd,J＝11.0,5.7Hz,1H),3.40(ddd,J＝7.0,5.8,2.9Hz,1H),3.01(s,3H),2.98–2.91(m,1H),2.80(dd,J＝4.9,2.6Hz,1H).MS:m/z calcd for C₁₈H₁₈N₃O₂ 308.1；found 308.3,M+H+.

example 6: synthesis of intermediate (S) -5

Dissolving intermediate (R) -4(283.4mg,0.9mmol) in 5mL of anhydrous tetrahydrofuran, gradually dropwise adding tetrabutylammonium fluoride (10.0mL,10.0mmol) after completion of the reaction, heating in a 90 ℃ oil bath for reflux reaction for 6 hours, removing the tetrahydrofuran by reduced pressure distillation after the reaction is completed, and performing column chromatography by using a developing solvent with the volume ratio of petroleum ether to ethyl acetate being 2:1 to obtain 230.6mg of yellow solid. Yield 76.2%, structure as follows:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.08(d,J＝8.0Hz,2H),7.88(d,J＝9.1Hz,1H),7.15(dd,J＝9.1,2.6Hz,1H),7.07(d,J＝8.9Hz,2H),7.01(d,J＝2.1Hz,1H),4.64(dt,J＝47.0,4.6Hz,2H),4.38–4.26(m,1H),4.21–4.12(m,1H),3.01(s,3H).HRMS:m/z calcd for C₁₈H₁₉FN₃O₂ 328.1461；found 328.1456,M+H+.

example 7: synthesis of intermediate (R) -5

Intermediate (R) -5 was prepared from intermediate (S) -4 according to the procedure for the synthesis of intermediate (S) -5 to give 183.4mg as a yellow solid. Yield 68.2%, structure is as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.08(d,J＝8.5Hz,2H),7.89(d,J＝8.9Hz,1H),7.16(d,J＝9.3Hz,1H),7.10–6.97(m,3H),4.63(dt,J＝47.1,5.2Hz,2H),4.31(d,J＝17.9Hz,1H),4.17(d,J＝4.9Hz,2H),3.02(s,3H).HRMS:m/z calcd for C₁₈H₁₉FN₃O₂ 328.1461；found 328.1456,M+H+.

example 8: synthesis of intermediate (R) -6

Intermediate 3(756.4mg,3.0mmol) and (R) - (-) -p-toluenesulphonic acid-2, 2-dimethyl-1, 3-dioxolanyl-4-methyl ester (1.0g,3.6mmol) were dissolved in 5mL of DMF, and cesium fluoride (1.4g,9.0mmol) was added under stirring and reacted at 65 ℃ for 6 hours. After the reaction is finished, after the reaction liquid is cooled to room temperature, 100mL of deionized water is added, a yellow product is separated out and is placed in ultrasound for 30 minutes, after suction filtration, the obtained solid is dissolved by dichloromethane and then is dissolved by anhydrous Na₂SO₄Drying, and separating by column chromatography with a developing solvent of petroleum ether and ethyl acetate in a volume ratio of 2:1 to obtain light yellow solid 1.0g, wherein the yield is 95.2%, and the structure is as follows:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.06(d,J＝7.6Hz,2H),7.84(d,J＝9.0Hz,1H),7.12(d,J＝9.2Hz,1H),7.09–7.02(m,2H),6.98(s,1H),4.58–4.46(m,1H),4.18(dd,J＝15.2,8.6Hz,1H),4.14(dd,J＝9.5,5.5Hz,1H),4.02(dd,J＝9.4,5.9Hz,1H),3.94(dd,J＝8.4,5.9Hz,1H),2.99(s,3H),1.48(s,3H),1.42(s,3H).MS:m/z calcd for C₂₁H₂₄N₃O₃366.2；found 366.4,M+H+.

example 9: synthesis of intermediate (S) -6

Intermediate (S) -6 was prepared from intermediate 3 according to the procedure for the synthesis of intermediate (R) -6 to give 1.0g of a pale yellow solid in 94.5% yield, having the following structure:¹H NMR(400MHz,CDCl₃)9.06(s,1H),8.19–7.95(m,2H),7.82(d,J＝9.0Hz,1H),7.15(d,J＝9.2Hz,1H),7.04(d,J＝8.7Hz,2H),6.98(s,1H),4.67–4.43(m,1H),4.19(dd,J＝8.5,6.4Hz,1H),4.11(d,J＝5.4Hz,1H),4.07–3.97(m,1H),3.93(dd,J＝8.5,5.9Hz,1H),2.98(s,3H),1.48(s,3H),1.42(s,3H).MS:m/z calcd for C₂₁H₂₄N₃O₃ 366.2；found 366.3,M+H+.

example 10: synthesis of intermediate (R) -7

Intermediate (R) -6(1.0g,2.8mmol) was dissolved in 10mL tetrahydrofuran, 1M hydrochloric acid was slowly added dropwise with stirring, and after the addition was completed, the reaction was refluxed in an oil bath at 90 ℃ for half an hour. After the reaction is finished, the tetrahydrofuran is removed by reduced pressure distillation, the pH value of the reaction solution is adjusted to be neutral, the reaction solution is placed in ultrasonic for 30 minutes, the product can be separated out, light yellow solid 910.2mg can be obtained by suction filtration and drying, the yield is 99.9%, and the structure is as follows:¹H NMR(400MHz,DMSO-d₆)9.21(s,1H),8.13(d,J＝8.6Hz,2H),7.73(d,J＝9.1Hz,1H),7.24(dd,J＝9.1,1.7Hz,1H),7.07(d,J＝8.6Hz,2H),6.72(s,1H),6.62(d,J＝4.8Hz,1H),4.96(d,J＝4.9Hz,1H),4.67(t,J＝5.4Hz,1H),4.07(dd,J＝9.8,4.0Hz,1H),3.93(dd,J＝9.7,6.2Hz,1H),3.87–3.66(m,1H),3.46(t,J＝5.3Hz,2H),2.81(d,J＝4.8Hz,3H).MS:m/z calcd for C₁₈H₂₀N₃O₃ 326.1；found 326.4,M+H+.

example 11: synthesis of intermediate (S) -7

Intermediate (S) -7 was prepared from intermediate (S) -6 according to the procedure for the synthesis of intermediate (R) -7 to give 1.1g of a pale yellow solid in 99.7% yield, the structure was as follows:¹H NMR(400MHz,DMSO-d₆)9.21(s,1H),8.13(d,J＝8.8Hz,2H),7.73(d,J＝9.1Hz,1H),7.24(dd,J＝9.1,2.5Hz,1H),7.07(d,J＝8.9Hz,2H),6.72(d,J＝2.4Hz,1H),6.62(q,J＝4.7Hz,1H),4.96(d,J＝5.2Hz,1H),4.66(t,J＝5.7Hz,1H),4.07(dd,J＝9.9,4.1Hz,1H),3.93(dd,J＝9.9,6.2Hz,1H),3.81(td,J＝12.2,5.6Hz,1H),3.45(t,J＝5.7Hz,2H),2.81(d,J＝4.9Hz,3H).MS:m/z calcd for C₁₈H₂₀N₃O₃ 326.1；found 326.3,M+H+.

example 12: synthesis of intermediate (S) -8

Intermediate (R) -7(0.6g,2.0mmol), t-butyldimethylsilyl chloride (1.8g,12.0mmol) and imidazole (1.1g,16.0mmol) were dissolved in 100mL of acetonitrile, stirred at room temperature for 12 hours, after completion of the reaction, the acetonitrile was distilled off under reduced pressure, and the reaction was continuedAdding dichloromethane, precipitating white powder, filtering to obtain lower layer filtrate, distilling under reduced pressure to remove dichloromethane, and separating by column chromatography with developing solvent (volume ratio of petroleum ether to ethyl acetate: 1) to obtain yellow solid 1.0g, with yield 93.7%, and structure as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.05(d,J＝8.6Hz,2H),7.88(d,J＝9.0Hz,1H),7.15(d,J＝9.1Hz,1H),7.09–6.89(m,3H),4.18(dd,J＝9.5,3.3Hz,1H),4.14–4.02(m,1H),3.95(dd,J＝9.4,6.8Hz,1H),3.67–3.65(dd,J＝5.6,2.9Hz,2H),3.01(s,3H),0.91(d,J＝1.5Hz,18H),0.12(d,J＝2.6Hz,6H),0.08(s,6H).MS:m/z calcd for C₃₀H₄₈N₃O₃Si₂ 554.3；found 554.7,M+H+.

example 13: synthesis of intermediate (R) -8

Intermediate (R) -8 was prepared from intermediate (S) -7 according to the procedure for the synthesis of intermediate (S) -8 to give 1.0g of a pale yellow solid in 87.2% yield, the structure was as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.05(d,J＝8.9Hz,2H),7.89(d,J＝9.1Hz,1H),7.15(dd,J＝9.1,2.5Hz,1H),7.05(d,J＝8.9Hz,3H),4.19(dd,J＝9.5,3.6Hz,1H),4.09(dt,J＝10.2,6.6Hz,1H),3.95(dd,J＝9.5,6.6Hz,1H),3.67(dd,J＝5.9,3.2Hz,2H),3.02(s,3H),0.91(d,J＝2.3Hz,18H),0.12(d,J＝3.0Hz,6H),0.08(d,J＝0.7Hz,6H).MS:m/z calcd for C₃₀H₄₈N₃O₃Si₂ 554.3；found 554.6,M+H+.

example 14: synthesis of intermediate (S) -9

Intermediate (S) -8(1.0g,2.0mmol) and di-tert-butyl dicarbonate (4.4g,20.0mmol) were dissolved in 30mL of tetrahydrofuran and heated in an oil bath at 90 ℃ for reflux reaction for 48 hours. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the light yellow solid is obtained by column chromatography separation by using a developing solvent with the volume ratio of petroleum ether to ethyl acetate being 5:1, wherein the light yellow solid is 1.0g, the yield is 80.6%, and the structure is as follows:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.16(d,J＝8.5Hz,2H),8.10(d,J＝9.1Hz,1H),7.99–7.64(m,2H),7.09(d,J＝8.5Hz,2H),4.21(dd,J＝9.4,3.1Hz,1H),4.10(s,1H),3.97(dd,J＝8.9,7.2Hz,1H),3.76–3.46(m,2H),3.44(s,3H),1.51(s,9H),0.91(d,J＝1.5Hz,18H),0.12(d,J＝3.1Hz,6H),0.08(s,6H).MS:m/z calcd for C₃₅H₅₆N₃O₅Si₂ 654.4；found 654.5,M+H+.

example 15: synthesis of intermediate (R) -9

Intermediate (R) -9 was prepared from intermediate (R) -8 according to the procedure for the synthesis of intermediate (S) -9 to give 1.2g of a pale yellow solid in 89.7% yield, the structure was as follows:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.17–8.10(m,3H),7.92–7.87(m,2H),7.09(d,J＝8.7Hz,1H),4.21(dd,J＝9.6,3.4Hz,1H),4.15–4.05(m,1H),3.97(dd,J＝9.5,6.7Hz,1H),3.67(t,J＝5.6Hz,2H),3.44(s,3H),1.51(s,9H),0.91(d,J＝2.5Hz,18H),0.12(d,J＝3.4Hz,6H),0.08(d,J＝0.7Hz,6H).MS:m/z calcd for C₃₅H₅₆N₃O₅Si₂ 654.4；found 654.6,M+H+.

example 16: synthesis of intermediate (R) -10

Intermediate (S) -9(1.0g,1.5mmol) and tetrabutylammonium fluoride (4.5mL,4.5mmol) were dissolved in 10mL tetrahydrofuran and heated in an oil bath at 90 ℃ for reflux for 3 hours. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the yellow solid is obtained by column chromatography separation by using a developing solvent with the volume ratio of petroleum ether to ethyl acetate being 1:4, wherein the yellow solid is 0.5g, the yield is 73.6%, and the structure is as follows:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.17(d,J＝7.6Hz,2H),8.09(dd,J＝9.0,3.8Hz,1H),7.91(s,2H),7.10(d,J＝8.1Hz,2H),4.21–4.10(m,3H),3.89(dd,J＝11.4,3.5Hz,1H),3.80(dd,J＝11.4,5.0Hz,1H),3.44(s,3H),1.51(s,9H).MS:m/z calcd for C₂₃H₂₈N₃O₅ 426.2；found 426.3,M+H+.

example 17: synthesis of intermediate (S) -10

Intermediate (S) -10 was prepared from intermediate (R) -9 according to the procedure for the synthesis of intermediate (R) -10 to give 0.6g of a yellow solid in 91.1% yield and the structure was as follows:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.17(d,J＝8.9Hz,2H),8.11(d,J＝9.0Hz,1H),7.99–7.84(m,2H),7.10(d,J＝8.9Hz,2H),4.16(d,J＝2.8Hz,3H),3.89(dd,J＝11.4,3.6Hz,1H),3.80(dd,J＝11.4,5.0Hz,1H),3.45(s,3H),1.51(s,9H).MS:m/z calcd for C₂₃H₂₈N₃O₅ 426.2；found 426.4,M+H+.

example 18: synthesis of intermediate (S) -11

Intermediate (R) -10(0.4g,1.0mmol) was dissolved in dichloromethane and p-methylbenzenesulfonyl chloride (0.2g,1.0mmol) was added slowly with stirring, then 5mL of triethylamine were added slowly and the reaction was stirred at room temperature for 6 hours. After the reaction is finished, the solvent is removed by distillation under the reduced pressure, and the yellow oily compound 163.7mg is obtained by column chromatography separation by using a developing solvent with the volume ratio of petroleum ether to ethyl acetate being 1:1, the yield is 27.8%, and the structure is as follows:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.16(d,J＝8.7Hz,2H),8.09(d,J＝9.0Hz,1H),7.88(d,J＝15.8Hz,2H),7.81(d,J＝8.3Hz,2H),7.34(d,J＝8.2Hz,2H),7.02(d,J＝8.7Hz,2H),4.40–4.18(m,3H),4.10(d,J＝4.5Hz,2H),3.44(s,3H),2.43(s,3H),1.51(s,9H).MS:m/z calcd for C₃₀H₃₄N₃O₇S 580.2；found 580.4,M+H+.

example 19: synthesis of intermediate (R) -11

Intermediate (R) -11 was prepared from intermediate (S) -10 according to the procedure for the synthesis of intermediate (S) -11 to give 215.3mg of compound as a yellow oil in 32.9% yield, having the following structure:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.16(d,J＝7.6Hz,2H),8.10(s,1H),7.91(s,2H),7.82(d,J＝8.3Hz,2H),7.34(d,J＝8.4Hz,2H),7.02(d,J＝7.6Hz,2H),4.48–4.20(m,3H),4.10(d,J＝4.4Hz,2H),3.45(s,3H),2.43(s,3H),1.51(s,9H).MS:m/z calcd for C₃₀H₃₄N₃O₇S 580.2；found 580.6,M+H+.

example 20: synthesis of intermediate (S) -12

Intermediate (S) -11(60.4mg,0.1mmol), dihydropyran (80.5mg,1.0mmol) and pyridinium 4-methylbenzenesulfonate (52.1mg,0.2mmol) were dissolved in anhydrous dichloromethane and stirred at room temperature for 5 hours. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the light yellow oily compound is obtained by column chromatography separation by using a developing solvent with the volume ratio of petroleum ether to ethyl acetate being 2:1, wherein the light yellow oily compound is 60.5mg, the yield is 90.4%, and the structure is as follows:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.16–8.10(m,3H),7.90(d,J＝12.6Hz,2H),7.79(dd,J＝8.2,5.8Hz,2H),7.30(d,J＝7.1Hz,2H),7.08–6.93(m,2H),4.36–4.17(m,3H),4.13–4.06(m,1H),3.92–3.80(m,2H),3.51–3.49(m,2H)3.45(d,J＝1.3Hz,3H),2.41(s,3H),1.82–1.67(m,2H),1.53–1.51(m,4H),1.51(d,J＝0.8Hz,9H).MS:m/z calcd for C₃₅H₄₂N₃O₈S 664.3；found 664.6,M+H+.

example 21: synthesis of intermediate (R) -12

Intermediate (2R) -12 was prepared from intermediate (R) -11 according to the procedure for the synthesis of intermediate (2S) -12 to give 0.1g of the compound as a pale yellow oil in 72.4% yield, which had the following structure:¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.35–8.07(m,3H),7.93(d,J＝11.6Hz,2H),7.80(dd,J＝11.5,8.0Hz,2H),7.32(dd,J＝14.0,8.1Hz,2H),7.08–6.86(m,2H),4.38–3.99(m,5H),3.90–3.70(m,2H),3.49(s,1H),3.45(s,3H),2.41(s,3H),1.78–1.70(m,4H),1.52(s,2H),1.52(s,9H).MS:m/z calcd for C₃₅H₄₂N₃O₈S 664.3；found 664.6,M+H+.

the synthetic procedures for the compounds of examples 1-21 above are schematically shown in FIG. 1.

Example 22: synthesis of intermediate 13

Intermediate 5-chloro-2-nitroaniline (12.5g,72.5mmol) and 50mL of dimethylamine were dissolved in 50mL of dimethylformamide, potassium carbonate was slowly added with stirring, and the reaction was heated in an oil bath at 90 ℃ under reflux for 12 hours. After the reaction is finished, cooling the reaction to room temperature, adding 200mL of cooled deionized water, placing the mixture in ultrasound for half an hour, performing suction filtration to collect solid, and drying the solid to obtain 8.0g of yellow solid, wherein the yield is 60.9 percent, and the structure is as follows:¹H NMR(400MHz,CDCl₃)8.02(d,J＝9.7Hz,1H),6.15(dd,J＝9.7,2.6Hz,1H),5.78(d,J＝2.6Hz,1H),3.05(s,6H).MS:m/z calcd for C₈H₁₂N₃O₂182.1；found 182.2,M+H+.

example 23: synthesis of intermediate 14

Dissolving the intermediate 13(3.6g,20.0mmol) and palladium carbon (1.0g,20.0mmol) in 130mL of methanol, continuously introducing hydrogen into the reaction system under stirring at room temperature for 6 hours, filtering to remove the palladium carbon, distilling under reduced pressure to remove the solvent, adding 5mL of dimethyl sulfoxide, slowly dropwise adding 2-bromo-4' -hydroxy dissolved in 5mL of dimethyl sulfoxide into the reaction solution in a water bath deviceAcetophenone (4.3g,20.0mmol) was added dropwise, the water bath was removed, and the reaction was stirred at room temperature for 10 hours. After the reaction was completed, the mixture was extracted 10 times with 50mL of ethyl acetate. The organic phase was dried over anhydrous magnesium sulfate and separated by column chromatography using a developing solvent in a volume ratio of methanol to dichloromethane of 5:1 to give 2.0g of a yellow solid which was a mixture of intermediate 14 and intermediate 21. Heating the mixture of the intermediate 14 and the intermediate 21 and oxalic acid dihydrate according to the ratio of 1:2, dissolving in ethanol with a small volume as far as possible to obtain a mauve solution, cooling, precipitating a mauve oxalate crystal of the intermediate 21, performing suction filtration, washing, adding a small amount of ammonia water to obtain a yellow solid, performing suction filtration, washing and drying to obtain the intermediate 21. Adding a large amount of petroleum ether into the mother liquor, precipitating dark red oxalate crystals of the intermediate 14, performing suction filtration, washing, adding a small amount of ammonia water to obtain yellow solid, performing suction filtration, washing and drying to obtain the intermediate 14. Yield 30.1%, structure as follows:¹H NMR(400MHz,DMSO-d₆)9.05(s,1H),8.12(d,J＝8.4Hz,2H),7.80(d,J＝9.3Hz,1H),7.41(dd,J＝9.2,1.9Hz,1H),6.93(dd,J＝12.6,5.1Hz,3H),3.08(s,6H).MS:m/z calcd for C₁₆H₁₆N₃O 266.1；found 266.2,M+H+.

example 24: synthesis of intermediate (R) -15

Intermediate (R) -15 was obtained from intermediate 14 according to the method for synthesizing intermediate (R) -4 to give 536.1mg as a pale yellow solid. Yield 83.4%, structure as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.12(d,J＝8.8Hz,2H),7.90(d,J＝9.3Hz,1H),7.33(dd,J＝9.3,2.8Hz,1H),7.15(s,1H),7.08(dd,J＝6.9,4.9Hz,2H),4.32(dd,J＝11.0,3.1Hz,1H),4.04(dd,J＝11.0,5.7Hz,1H),3.44–3.36(m,1H),3.15(s,6H),2.97–2.91(m,1H),2.80(dd,J＝4.9,2.6Hz,1H).MS:m/z calcd for C₁₉H₂₀N₃O₂ 322.1；found 322.2,M+H+.

example 25: synthesis of intermediate (S) -15

Intermediate (S) -15 was obtained from intermediate 14 according to the procedure for the synthesis of intermediate (R) -4 to give 581.2mg as a pale yellow solid. Yield 90.7%, structure is as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.12(d,J＝8.8Hz,2H),7.89(d,J＝9.3Hz,1H),7.33(dd,J＝9.3,2.8Hz,1H),7.13(s,1H),7.10–7.03(m,2H),4.32(dd,J＝11.0,3.1Hz,1H),4.04(dd,J＝11.0,5.7Hz,1H),3.44–3.36(m,1H),3.15(s,6H),2.95–2.88(m,1H),2.80(dd,J＝4.9,2.6Hz,1H).MS:m/z calcd for C₁₉H₂₀N₃O₂ 322.1；found 322.3,M+H+.

example 26: synthesis of intermediate (S) -16

Intermediate (S) -16 was prepared from intermediate (R) -15 according to the procedure for the synthesis of intermediate (S) -5 to give 196.5mg of a yellow solid. Yield 84.6%, structure as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.14(d,J＝8.8Hz,2H),7.92(d,J＝9.3Hz,1H),7.35(dd,J＝9.3,2.8Hz,1H),7.20(s,1H),7.08(d,J＝8.9Hz,2H),4.63(dt,J＝47.0,4.7Hz,2H),4.33-4.28(m,1H),4.18–4.16(dd,J＝4.6,2.6Hz,2H),3.17(s,6H).HRMS:m/z calcd for C₁₉H₂₁FN₃O₂342.1612；found 342.1615,M+H+.

example 27: synthesis of intermediate (R) -16

Intermediate (R) -16 was prepared from intermediate (S) -15 according to the procedure for the synthesis of intermediate (S) -5 to give 158.3mg as a yellow solid. Yield 86.7%, structure is as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.18(d,J＝8.2Hz,2H),7.94(d,J＝9.4Hz,1H),7.37(dd,J＝9.4,2.4Hz,1H),7.10(d,J＝8.9Hz,2H),4.63(dt,J＝47.1,5.0Hz,2H),4.34-4.29(m,1H),4.18-4.17(m,2H),3.19(s,6H).HRMS:m/z calcd for C₁₉H₂₁FN₃O₂ 342.1612；found 342.1609,M+H+.

example 28: synthesis of intermediate (R) -17

Intermediate (R) -17 was prepared from intermediate 14 according to the procedure for the synthesis of intermediate (R) -6 to give 756.0mg as a pale yellow solid in 99.6% yield, having the following structure:¹H NMR(400MHz,CDCl₃)8.93(s,1H),8.13(d,J＝3.2Hz,2H),7.92(s,1H),7.35(s,1H),7.18(s,1H),7.08(s,2H),4.69–4.40(m,1H),4.20(dd,J＝8.5,6.4Hz,1H),4.14(dd,J＝9.5,5.5Hz,1H),4.05(d,J＝5.8Hz,1H),3.94(dd,J＝8.5,5.9Hz,1H),3.17(s,6H),1.48(s,3H),1.32(s,3H).MS:m/z calcd for C₂2H₂₆N₃O₃ 380.2；found 380.4,M+H+.

example 29: synthesis of intermediate (S) -17

Intermediate (S) -17 was prepared from intermediate 14 according to the procedure for the synthesis of intermediate (R) -6 to give 630.2mg as a pale yellow solid in 83.4% yield, having the following structure:¹H NMR(400MHz,CDCl₃)8.93(s,1H),8.13(d,J＝8.7Hz,2H),7.92(d,J＝9.3Hz,1H),7.35(dd,J＝9.4,2.4Hz,1H),7.09(d,J＝8.8Hz,2H),4.51(dd,J＝11.8,5.9Hz,1H),4.20(dd,J＝8.5,6.5Hz,1H),4.14(dd,J＝9.6,5.5Hz,1H),4.04(dd,J＝9.5,5.8Hz,1H),3.93(dd,J＝8.5,5.9Hz,1H),3.18(s,6H),1.48(s,3H),1.42(s,3H).MS:m/z calcd for C₂₂H₂₆N_3O3 380.2；found 380.4,M+H+.

example 30: synthesis of intermediate (R) -18

Intermediate (R) -18 was prepared from intermediate (R) -17 according to the procedure for the synthesis of intermediate (R) -7 to give 676.3mg as a pale yellow solid, 98.4% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.10(s,1H),8.22(d,J＝8.5Hz,2H),7.82(d,J＝9.3Hz,1H),7.43(dd,J＝9.3,2.0Hz,1H),7.09(d,J＝8.5Hz,2H),6.96(d,J＝1.9Hz,1H),4.98(d,J＝5.0Hz,1H),4.69(t,J＝5.6Hz,1H),4.08(dd,J＝9.8,4.0Hz,1H),3.94(dd,J＝9.8,6.2Hz,1H),3.82(dd,J＝10.1,5.2Hz,1H),3.46(t,J＝5.6Hz,2H),3.09(s,6H).MS:m/z calcd for C₁₉H₂₂N₃O₃ 340.2；found 340.4,M+H+.

example 31: synthesis of intermediate (S) -18

Intermediate (S) -18 was prepared from intermediate (S) -17 according to the procedure for the synthesis of intermediate (R) -7 to give 500.4mg as a pale yellow solid in 88.5% yield, having the structure:¹H NMR(400MHz,DMSO-d₆)9.28(s,1H),8.19(dd,J＝19.8,8.5Hz,2H),7.84(dd,J＝14.4,9.3Hz,1H),7.47(dd,J＝26.6,9.2Hz,1H),7.09(t,J＝7.7Hz,2H),6.97(s,1H),4.98(s,1H),4.68(s,2H),4.09–4.05(m,1H),4.02–3.88(m,1H),3.81(s,1H),3.46(s,2H),3.09(s,6H).MS:m/z calcd for C₁₉H₂₂N₃O₃ 340.2；found 340.5,M+H+.

example 32: synthesis of intermediate (S) -19

From the intermediate (R) -11 according to the method for synthesizing the intermediate (S) -11Intermediate (S) -19 was prepared 18 to give 260.5mg of compound as a yellow oil in 38.2% yield, the structure is as follows:¹H NMR(400MHz,CDCl₃)8.91(s,1H),8.07(d,J＝8.9Hz,2H),7.90(d,J＝9.3Hz,1H),7.79(d,J＝8.3Hz,1H),7.32(dd,J＝13.3,5.5Hz,3H),7.10(d,J＝2.6Hz,1H),7.07–7.02(m,1H),6.94(dd,J＝8.8,3.9Hz,1H),4.33–4.18(m,3H),4.12(d,J＝7.2Hz,1H),4.05(d,J＝4.5Hz,1H),3.15(s,6H),2.40(s,3H).MS:m/z calcd for C₂₆H₂₈N₃O₅S 494.2；found 494.5,M+H+.

example 33: synthesis of intermediate (R) -19

Intermediate (R) -19 was prepared from intermediate (S) -18 according to the procedure for the synthesis of intermediate (S) -11 to give 170.7mg of compound as a yellow oil in 33.8% yield, having the following structure:¹H NMR(400MHz,CDCl₃)8.92(s,1H),8.21–7.99(m,2H),7.90(d,J＝9.3Hz,1H),7.80(d,J＝8.3Hz,2H),7.33(dd,J＝13.3,5.4Hz,3H),7.11(dd,J＝13.4,2.5Hz,1H),6.98–6.89(m,2H),4.26(ddd,J＝16.0,11.3,5.5Hz,3H),4.13–3.97(m,2H),3.16(s,6H),2.41(s,3H).MS:m/z calcd for C₂₆H₂₈N₃O₅S 494.2；found 494.7,M+H+.

example 34: synthesis of intermediate (S) -20

Intermediate (2S) -20 was prepared from intermediate (S) -19 according to the procedure for the synthesis of intermediate (2S) -12 to give 120.8mg of the compound as a pale yellow oil in 77.5% yield, which had the following structure:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.11(d,J＝8.7Hz,2H),7.91(d,J＝9.3Hz,1H),7.79(dd,J＝8.2,5.6Hz,2H),7.35(dd,J＝9.3,2.7Hz,1H),7.32–7.27(m,1H),7.22(s,1H),7.08(dd,J＝11.7,5.2Hz,1H),6.96(dd,J＝8.8,5.8Hz,1H),4.36–4.05(m,6H),3.57–3.47(m,2H),3.17(s,6H),2.40(s,3H),1.82–1.67(m,2H),1.56–1.50(m,4H).MS:m/z calcd for C₃₁H₃6N₃O₆S 578.2；found578.6,M+H+.

example 35: synthesis of intermediate (R) -20

Intermediate (2R) -20 was prepared from intermediate (R) -19 according to the procedure for the synthesis of intermediate (2S) -12 to give 62.1mg of the compound as a pale yellow oil in 74.4% yield, which had the following structure:¹H NMR(400MHz,CDCl₃)8.93(s,1H),8.21(s,2H),8.00(dd,J＝25.9,9.2Hz,2H),7.79(dd,J＝8.2,5.8Hz,2H),7.40(d,J＝7.6Hz,1H),7.31(d,J＝7.0Hz,2H),7.06–6.88(m,2H),4.40–4.14(m,4H),4.12–4.06(m,2H),3.50(s,2H),3.23(s,6H),2.42(s,3H),1.53(s,4H),1.26(s,2H).MS:m/z calcd for C₃₁H₃₆N₃O₆S 578.2；found 578.6,M+H+.

example 36: synthesis of intermediate 21

Intermediate 21 was prepared from intermediate 13 according to the procedure for the synthesis of intermediate 14, in 6.3% yield, and the structure is as follows:¹H NMR(400MHz,DMSO-d₆)9.21(s,1H),8.04(d,J＝8.4Hz,2H),7.81(d,J＝9.3Hz,1H),7.48(dd,J＝9.2,1.9Hz,1H),6.95-6.86(m,3H),3.06(s,6H).MS:m/z calcd for C₁₆H₁₆N₃O 266.1；found 266.2,M+H+.

example 37: synthesis of intermediate (R) -22

Intermediate (R) -22 was obtained from intermediate 21 according to the method for synthesizing intermediate (R) -4 to give 536.1mg as a pale yellow solid. Yield 83.4%, structure as follows:¹H NMR(400MHz,CDCl₃)9.09(s,1H),8.21–8.01(m,2H),7.94(d,J＝9.3Hz,1H),7.39(dd,J＝9.4,2.8Hz,1H),7.09(d,J＝2.8Hz,1H),7.06(d,J＝1.9Hz,2H),4.31(dd,J＝11.0,3.1Hz,1H),4.04(dd,J＝11.0,5.7Hz,1H),3.40(ddd,J＝7.0,5.8,3.0Hz,1H),3.15(s,6H),3.00–2.89(m,1H),2.80(dd,J＝4.9,2.6Hz,1H).MS:m/z calcd for C₁₉H₂₀N₃O₂ 322.1；found 322.3,M+H+.

example 38: synthesis of intermediate (S) -22

Intermediate (S) -22 was obtained from intermediate 21 according to the method for synthesizing intermediate (R) -4 to give 532.5mg as a pale yellow solid. Yield 84.9%, structure as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.08(d,J＝8.9Hz,2H),7.96(d,J＝9.4Hz,1H),7.41(dd,J＝9.4,2.7Hz,1H),7.13(d,J＝2.5Hz,1H),7.07(d,J＝8.8Hz,2H),4.32(dd,J＝11.0,3.1Hz,1H),4.04(dd,J＝11.0,5.7Hz,1H),3.40(dt,J＝8.6,3.0Hz,1H),3.17(s,6H),2.94(t,J＝4.5Hz,1H),2.80(dd,J＝4.9,2.6Hz,1H).MS:m/z calcd for C₁₉H₂₀N₃O₂ 322.1；found 322.3,M+H+.

example 39: synthesis of intermediate (S) -23

Intermediate (S) -23 was prepared from intermediate (R) -22 according to the procedure for the synthesis of intermediate (S) -5 to give 187.2mg as a yellow solid. Yield 85.3%, structure as follows:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.08(d,J＝8.8Hz,2H),7.95(d,J＝9.3Hz,1H),7.41(dd,J＝9.4,2.7Hz,1H),7.12(s,1H),7.07(d,J＝8.8Hz,2H),4.75–4.64(m,1H),4.62–4.50(m,1H),4.41–4.23(m,1H),4.16(d,J＝4.8Hz,2H),3.16(s,6H).HRMS:m/z calcd for C₁₉H₂₁FN₃O₂342.1618；found,M+H+.

example 40: synthesis of intermediate (R) -23

Intermediate (S) -23 was prepared from intermediate (S) -22 according to the procedure for the synthesis of intermediate (S) -5 to give 168.2mg as a yellow solid. Yield 82.5%, structure is as follows:¹H NMR(400MHz,CDCl₃)9.09(s,1H),8.08(dd,J＝9.0,2.4Hz,2H),7.94(d,J＝9.3Hz,1H),7.39(dd,J＝9.4,2.6Hz,1H),7.07(dt,J＝9.1,2.6Hz,3H),4.76–4.63(m,1H),4.62–4.51(m,1H),4.38–4.25(m,1H),4.16(dd,J＝4.0,1.7Hz,2H),3.15(s,6H).HRMS:m/z calcd for C₁₉H₂₁FN₃O₂342.1618；found,M+H+.

example 41: synthesis of intermediate (R) -24

Intermediate (R) -24 was prepared from intermediate 21 according to the procedure for the synthesis of intermediate (R) -6 to give 673.1mg as a pale yellow solid in 93.5% yield, and the structure was as follows:¹H NMR(400MHz,CDCl₃)9.09(s,1H),8.12–8.04(m,2H),7.94(d,J＝9.3Hz,1H),7.39(d,J＝9.3Hz,1H),7.10–7.05(m,3H),4.52(p,J＝5.8Hz,1H),4.20(dd,J＝8.5,6.4Hz,1H),4.14(dd,J＝9.5,5.5Hz,1H),4.03(dd,J＝9.5,5.8Hz,1H),3.94(dd,J＝8.5,5.9Hz,1H),3.16(d,J＝3.0Hz,6H),1.49(s,3H),1.42(s,3H).MS:m/z calcd for C₂₂H₂₆N₃O₃ 380.2；found 380.4,M+H+.

example 42: synthesis of intermediate (S) -24

Intermediate (S) -24 was prepared from intermediate 21 according to the procedure for the synthesis of intermediate (R) -6 to give 660.1mg as a pale yellow solid in 91.4% yield, the structure of which is as follows:¹H NMR(400MHz,CDCl₃)9.09(s,1H),8.07(d,J＝8.9Hz,2H),7.95(d,J＝9.3Hz,1H),7.48–7.30(m,1H),7.11(s,1H),7.07(t,J＝5.9Hz,2H),4.59–4.40(m,1H),4.20(dd,J＝8.5,6.4Hz,1H),4.14(dd,J＝9.5,5.5Hz,1H),4.03(dd,J＝9.5,5.8Hz,1H),3.94(dd,J＝8.5,5.9Hz,1H),3.16(s,6H),1.49(s,3H),1.42(s,3H).MS:m/z calcd for C₂₂H₂₆N₃O₃ 380.2；found 380.4,M+H+.

example 43: synthesis of intermediate (R) -25

Intermediate (R) -25 was prepared from intermediate (R) -24 according to the procedure for the synthesis of intermediate (R) -7 to give 594.2mg as a pale yellow solid in 95.6% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.27(s,1H),8.17(d,J＝8.9Hz,1H),7.85(d,J＝9.3Hz,1H),7.50(dd,J＝9.4,2.8Hz,1H),7.08(d,J＝8.9Hz,2H),6.97(d,J＝2.8Hz,1H),4.98(d,J＝4.8Hz,1H),4.68(s,1H),4.07(dd,J＝9.9,4.1Hz,1H),3.93(dd,J＝9.9,6.2Hz,1H),3.81(d,J＝5.0Hz,1H),3.45(t,J＝4.8Hz,2H),3.09(s,6H).MS:m/z calcd for C₁₉H₂₂N₃O₃ 340.2；found 340.3,M+H+.

example 44: synthesis of intermediate (S) -25

Intermediate (S) -25 was prepared from intermediate (S) -24 according to the procedure for the synthesis of intermediate (R) -7 to give 578.4mg as a pale yellow solid in 94.1% yield, the structure is as follows:¹H NMR(400MHz,DMSO-d₆)9.27(s,1H),8.17(d,J＝8.9Hz,2H),7.85(d,J＝9.3Hz,1H),7.50(dd,J＝9.4,2.8Hz,1H),7.08(d,J＝8.9Hz,2H),6.97(d,J＝2.8Hz,1H),4.97(s,1H),4.68(s,1H),4.07(dd,J＝9.9,4.1Hz,1H),3.93(dd,J＝9.9,6.2Hz,1H),3.82(dd,J＝10.0,5.5Hz,1H),3.45(d,J＝5.7Hz,2H),3.09(s,6H).MS:m/z calcd for C₁₉H₂₂N₃O₃ 340.2；found 340.4,M+H+.

example 45: synthesis of intermediate (S) -26

Intermediate (S) -26 was prepared from intermediate (R) -25 according to the procedure for the synthesis of intermediate (S) -11 to give 265.2mg of compound as a yellow oil in 37.9% yield, having the following structure:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.06(d,J＝8.8Hz,2H),7.95(d,J＝9.3Hz,1H),7.80(d,J＝8.3Hz,2H),7.41(d,J＝9.4Hz,1H),7.32(d,J＝8.0Hz,2H),7.11(s,1H),6.96(d,J＝8.6Hz,2H),4.39–4.12(m,3H),4.07(d,J＝4.5Hz,2H),3.16(s,6H),2.42(s,3H).MS:m/z calcd for C₂₆H₂₈N₃O₅S 494.2；found 494.4,M+H+.

example 46: synthesis of intermediate (R) -26

Intermediate (R) -26 was prepared from intermediate (S) -25 according to the procedure for the synthesis of intermediate (S) -11 to give 193.6mg of compound as a yellow oil in 35.9% yield, the structure of which was as follows:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.06(d,J＝8.8Hz,2H),7.95(d,J＝9.4Hz,1H),7.81(d,J＝8.3Hz,2H),7.41(dd,J＝9.4,2.7Hz,1H),7.33(d,J＝8.3Hz,2H),7.12(s,1H),6.98(t,J＝7.0Hz,2H),4.24(ddd,J＝19.4,11.6,7.7Hz,3H),4.07(d,J＝4.5Hz,2H),3.17(s,6H),2.42(s,3H).MS:m/z calcd for C₂₆H₂₈N₃O₅S 494.2；found 494.4,M+H+.

example 47: synthesis of intermediate (S) -27

Intermediate (2S) -27 was prepared from intermediate (S) -26 according to the procedure for the synthesis of intermediate (2S) -12 to give 127.4mg of the compound as a pale yellow oil in 87.5% yield, which had the following structure:¹H NMR(400MHz,CDCl₃)9.06(s,1H),8.05(d,J＝8.3Hz,2H),7.97(d,J＝9.3Hz,1H),7.79(dd,J＝8.2,5.7Hz,2H),7.43(dd,J＝9.4,2.6Hz,1H),7.29(d,J＝7.3Hz,2H),7.15(s,1H),6.95(dd,J＝8.8,5.7Hz,2H),4.42–4.13(m,4H),4.12–3.99(m,2H),3.52–3.46(dd,J＝14.1,9.0Hz,2H),3.18(s,6H),2.40(s,3H),1.82–1.59(m,2H),1.53(s,4H).MS:m/z calcd for C₃₁H₃₆N₃O₆S 578.2；found 578.5,M+H+.

a schematic of the synthesis of the compounds of examples 22-48 above is shown in FIG. 2.

Example 48: synthesis of intermediate (R) -27

Intermediate (2R) -27 was prepared from intermediate (R) -26 according to the procedure for the synthesis of intermediate (2S) -12 to give 100.7mg of the compound as a pale yellow oil in 90.4% yield, which had the following structure:¹H NMR(400MHz,CDCl₃)9.05(s,1H),8.04(d,J＝8.3Hz,2H),7.98(d,J＝9.1Hz,1H),7.84–7.74(m,2H),7.45(d,J＝7.6Hz,1H),7.29(d,J＝7.9Hz,2H),7.18(s,1H),6.95(dd,J＝8.8,5.6Hz,2H),4.36–4.14(m,4H),4.13–4.01(m,2H),3.49(s,2H),3.19(s,6H),2.41(s,3H),1.92–1.64(m,2H),1.53(s,4H).MS:m/z calcd for C₃₁H₃₆N₃O₆S 578.2；found 578.4,M+H+.

example 49: synthesis of intermediate (R, R) -28

Intermediate (R, R) -28 was prepared from intermediate 14 according to the procedure for the synthesis of intermediate (R) -6 to give 248.8mg as a pale yellow solid in 43.9% yield, and the structure was as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.13(d,J＝8.8Hz,2H),7.91(d,J＝9.3Hz,1H),7.81(d,J＝8.3Hz,2H),7.34(d,J＝8.0Hz,3H),7.20(s,1H),7.02(d,J＝8.8Hz,2H),4.38–4.15(m,5H),4.13–4.02(m,1H),3.17(s,6H),2.43(s,3H),1.43(s,3H),1.40(s,3H).MS:m/z calcd for C₃₀H₃₄N₃O₆S 564.2；found 564.4,M+H+.

example 50: synthesis of intermediate (S, S) -28

Intermediate (S, S) -28 was prepared from intermediate 14 according to the procedure for the synthesis of intermediate (R) -6 to give 280.6mg as a pale yellow solid in 49.6% yield, the structure of which is as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.13(d,J＝8.8Hz,2H),7.91(d,J＝9.3Hz,1H),7.81(d,J＝8.3Hz,2H),7.42–7.28(m,3H),7.17(d,J＝19.6Hz,1H),7.02(d,J＝8.8Hz,2H),4.23(ddd,J＝13.9,12.0,4.3Hz,5H),4.14–4.01(m,1H),3.17(s,6H),2.43(s,3H),1.43(s,3H),1.40(s,3H).MS:m/z calcd for C₃₀H₃₄N₃O₆S 564.2；found 564.4,M+H+.

example 51: synthesis of intermediate (R, S) -29

Intermediate (R, S) -29 was prepared from intermediate (R, R) -28 according to the procedure for the synthesis of intermediate (S) -5 to give 54.3mg as a yellow solid. Yield 94.0%, structure is as follows:¹H NMR(400MHz,CDCl₃)8.94(s,1H),8.12(d,J＝8.8Hz,2H),7.90(d,J＝9.3Hz,1H),7.33(dd,J＝9.3,2.8Hz,1H),7.11(d,J＝2.7Hz,1H),7.09–6.98(m,2H),4.74(dd,J＝10.2,3.3Hz,0.5H),4.68–4.60(m,1H),4.54(dd,J＝10.2,4.5Hz,0.5H),4.42–4.31(m,1H),4.26(dq,J＝8.0,4.3Hz,2H),4.20–4.11(m,1H),3.15(s,6H),1.49(d,J＝2.3Hz,6H).MS:m/z calcd for C₂₃H₂₇FN₃O₃ 412.2；found 412.4,M+H+.

example 52: synthesis of intermediate (S, R) -29

Intermediate (S, R) -29 was prepared from intermediate (S, S) -28 according to the procedure for the synthesis of intermediate (S) -5 to give 103.7mg as a yellow solid. Yield 83.2%, structure as follows:¹H NMR(400MHz,CDCl₃)8.93(s,1H),8.15–8.09(m,2H),7.89(d,J＝9.3Hz,1H),7.32(dd,J＝9.3,2.8Hz,1H),7.10(d,J＝2.8Hz,1H),7.09–6.98(m,2H),4.73(dd,J＝10.2,3.3Hz,0.5H),4.69–4.58(m,1H),4.53(dd,J＝10.2,4.5Hz,0.5H),4.34(dt,J＝8.1,5.1Hz,1H),4.30–4.20(m,2H),4.16(dd,J＝9.7,5.2Hz,1H),3.14(s,6H),1.49(d,J＝2.2Hz,6H).MS:m/z calcd for C₂₃H₂₇FN₃O₃ 412.2；found 412.4,M+H+.

example 53: synthesis of intermediate (R, S) -30

Intermediate (R, S) -30 was prepared from intermediate (R, S) -29 according to the procedure for the synthesis of intermediate (R) -7 to give 37.5mg as a pale yellow solid in 83.3% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.10(s,1H),8.22(d,J＝8.9Hz,2H),7.82(d,J＝9.3Hz,1H),7.44(dd,J＝9.3,2.8Hz,1H),7.10(d,J＝8.9Hz,2H),6.96(d,J＝2.8Hz,1H),5.15(dd,J＝11.8,5.9Hz,2H),4.59(dd,J＝9.3,3.9Hz,0.5H),4.53–4.43(m,1H),4.35(dd,J＝9.3,6.6Hz,0.5H),4.14(dd,J＝9.7,4.5Hz,1H),3.97(dd,J＝9.6,6.5Hz,1H),3.91–3.75(m,2H),3.09(s,6H).HRMS:m/z calcd for C₂₀H₂₃FN₃O₃372.1645；found,M+H+.

example 54: synthesis of intermediate (S, R) -30

Intermediate (S, R) -30 was prepared from intermediate (S, R) -29 according to the procedure for the synthesis of intermediate (R) -7 to give 86.4mg as a pale yellow solid in 96.7% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.10(s,1H),8.22(d,J＝8.9Hz,2H),7.82(d,J＝9.3Hz,1H),7.43(dd,J＝9.3,2.8Hz,1H),7.10(d,J＝8.9Hz,2H),6.96(d,J＝2.8Hz,1H),5.15(dd,J＝11.8,5.9Hz,2H),4.59(dd,J＝9.3,3.9Hz,0.5H),4.53–4.43(m,1H),4.35(dd,J＝9.3,6.6Hz,0.5H),4.14(dd,J＝9.7,4.5Hz,1H),3.98(dd,J＝9.6,6.5Hz,1H),3.87–3.82(m,2H),3.09(s,6H).HRMS:m/z calcd for C₂₀H₂₃FN₃O₃372.1645；found,M+H+.

example 55: synthesis of intermediate (R, R) -31

Intermediate (R, R) -31 was prepared from intermediate 21 according to the procedure for the synthesis of intermediate (R) -6 to give 140.4mg as a pale yellow solid in 24.8% yield, the structure was as follows:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.07(d,J＝8.8Hz,2H),7.96(d,J＝9.4Hz,1H),7.81(d,J＝8.3Hz,2H),7.42(dd,J＝9.4,2.7Hz,1H),7.34(d,J＝8.1Hz,2H),7.13(d,J＝2.5Hz,1H),7.00(d,J＝8.8Hz,2H),4.42–4.15(m,5H),4.09(dd,J＝9.3,4.5Hz,1H),3.17(s,6H),2.44(s,3H),1.43(s,3H),1.40(s,3H).MS:m/z calcd for C₃₀H₃₄N₃O₆S 564.2；found 564.4,M+H+.

example 56: synthesis of intermediate (S, S) -31

Intermediate (S, S) -31 was prepared from intermediate 21 according to the procedure for the synthesis of intermediate (R) -6 to give 150.3mg as a pale yellow solid in 53.2% yield, the structure of which is as follows:¹H NMR(400MHz,CDCl₃)9.09(s,1H),8.07(d,J＝8.8Hz,2H),7.95(d,J＝9.3Hz,1H),7.81(d,J＝8.3Hz,2H),7.40(d,J＝9.3Hz,1H),7.33(d,J＝8.1Hz,2H),7.10(s,1H),7.00(d,J＝8.8Hz,2H),4.34–4.13(m,5H),4.08(dd,J＝9.3,4.5Hz,1H),3.16(s,6H),2.43(s,3H),1.43(s,3H),1.40(s,3H).MS:m/z calcd for C₃₀H₃₄N₃O₆S 564.2；found 564.4,M+H+.

example 57: synthesis of intermediate (R, S) -32

Intermediate (R, S) -32 was prepared from intermediate (R, R) -31 according to the procedure for the synthesis of intermediate (S) -5 to give 61.2mg as a yellow solid. Yield 92.5%, structure as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.08(d,J＝8.9Hz,2H),7.96(d,J＝9.4Hz,1H),7.42(dd,J＝9.4,2.8Hz,1H),7.13(d,J＝2.7Hz,1H),7.07(d,J＝6.7Hz,2H),4.74(dd,J＝10.2,3.3Hz,0.5H),4.70–4.57(m,1H),4.54(dd,J＝10.1,4.5Hz,0.5H),4.31–4.01(m,4H),3.17(s,6H),1.49(d,J＝2.3Hz,6H).MS:m/z calcd for C₂₃H₂₇FN₃O₃ 412.2；found 412.4,M+H+.

example 58: synthesis of intermediate (S, R) -32

Intermediate (S, R) -32 was prepared from intermediate (S, S) -31 according to the procedure for the synthesis of intermediate (S) -5 to give 97.4mg as a yellow solid. The yield is 80.4%, and the structure is as follows:¹H NMR(400MHz,CDCl₃)9.10(s,1H),8.11–8.07(m,2H),7.94(d,J＝9.3Hz,1H),7.39(dd,J＝9.4,2.7Hz,1H),7.13–6.93(m,3H),4.73(dd,J＝10.2,3.3Hz,0.5H),4.69–4.59(m,1H),4.53(dd,J＝10.2,4.5Hz,0.5H),4.37–4.32(m,1H),4.30–4.23(m,2H),4.18–4.14(m,1H),3.15(s,6H).MS:m/z calcd for C₂₃H₂₇FN₃O₃ 412.2；found 412.4,M+H+.

example 59: synthesis of intermediate (R, S) -33

Intermediate (R, S) -33 was prepared from intermediate (R, S) -32 according to the procedure for the synthesis of intermediate (R) -7 to give 84.2mg as a pale yellow solid in 73.2% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.28(s,1H),8.17(d,J＝8.8Hz,2H),7.86(d,J＝9.3Hz,1H),7.50(dd,J＝9.4,2.7Hz,1H),7.08(d,J＝8.8Hz,2H),6.97(d,J＝2.6Hz,1H),5.15(dd,J＝12.7,5.9Hz,2H),4.58(dd,J＝9.1,3.8Hz,0.5H),4.47(dd,J＝9.3,4.3Hz,1H),4.44–4.27(m,0.5H),4.13(dd,J＝9.6,4.4Hz,1H),3.97(d,J＝6.5Hz,1H),3.87–3.81(m,2H),3.09(s,6H).HRMS:m/z calcd for C₂₀H₂₃FN₃O₃ 372.1645；found,M+H+.

example 60: synthesis of intermediate (S, R) -33

Intermediate (S, R) -33 was prepared from intermediate (S, R) -32 according to the procedure for the synthesis of intermediate (R) -7 to give 86.4mg as a pale yellow solid in 89.6% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.27(s,1H),8.17(d,J＝8.9Hz,2H),7.86(d,J＝9.3Hz,1H),7.50(dd,J＝9.4,2.8Hz,1H),7.08(d,J＝8.9Hz,2H),6.97(d,J＝2.8Hz,1H),5.13(dd,J＝12.3,5.5Hz,2H),4.59(dd,J＝9.3,3.9Hz,0.5H),4.56–4.44(m,1H),4.35(dd,J＝9.3,6.6Hz,0.5H),4.13(dd,J＝9.7,4.5Hz,1H),3.97(dd,J＝9.5,6.6Hz,1H),3.87–3.81(m,2H),3.09(s,6H).HRMS:m/z calcd for C₂₀H₂₃FN₃O₃372.1645；found,M+H+.

example 61: synthesis of intermediate (S, S) -34

Intermediate (S, S) -34 was prepared from intermediate 3 according to the procedure for the synthesis of intermediate (R) -6 to give 345.8mg as a pale yellow solid in 62.7% yield, the structure is as follows:¹H NMR(400MHz,CDCl₃)9.07(s,1H),8.07(s,2H),7.96–7.73(m,3H),7.41–7.24(m,2H),7.15(s,1H),7.00(s,3H),4.27–4.09(m,7H),3.00(s,3H),2.43(s,3H),1.41(d,J＝12.0Hz,6H).MS:m/z calcd for C₂₉H₃₂N₃O₆S 550.2；found 550.4,M+H+.

example 62: synthesis of intermediate (S, R) -35

Intermediate (S, R) -35 was prepared from intermediate (S, S) -34 according to the procedure for the synthesis of intermediate (S) -5 to give 201.5mg as a yellow solid. Yield 84.1%, structure as follows:¹H NMR(400MHz,CDCl₃)9.08(s,1H),8.19–7.98(m,2H),7.86(d,J＝9.0Hz,1H),7.13(d,J＝9.0Hz,1H),7.11–7.03(m,2H),6.99(s,1H),5.06–4.45(m,2H),4.44–3.90(m,4H),3.00(s,3H),1.49(s,6H).MS:m/z calcd for C₂₂H₂₅FN₃O₃398.2；found 398.4,M+H+.

example 63: synthesis of intermediate (R, S) -36

Intermediate (R, S) -36 was prepared from intermediate (S, R) -35 according to the procedure for the synthesis of intermediate (R) -7 to give 70.5mg as a pale yellow solid in 39.1% yield, having the following structure:¹H NMR(400MHz,DMSO-d₆)9.21(s,1H),8.14(d,J＝8.8Hz,2H),7.73(d,J＝9.1Hz,1H),7.24(dd,J＝9.1,2.5Hz,1H),7.07(d,J＝8.9Hz,2H),6.72(d,J＝2.4Hz,1H),6.62(q,J＝4.9Hz,1H),5.13(dd,J＝13.2,5.8Hz,2H),4.58(dd,J＝9.3,3.9Hz,0.5H),4.52–4.42(m,1H),4.35(dd,J＝9.3,6.6Hz,0.5H),4.13(dd,J＝9.7,4.5Hz,1H),3.96(dd,J＝9.5,6.5Hz,1H),3.93–3.71(m,2H),2.81(d,J＝4.9Hz,3H).HRMS:m/z calcd for C₁₉H₂₁FN₃O₃ 358.1489；found,M+H+.

a schematic of the synthesis of the compounds of examples 49-63 above is shown in FIG. 3.

Example 64: synthesis of 3- (4-nitrophenyl) quinoxalin-6-ol (37)

2-bromo-4' -nitroacetophenone (2.44g,10.0mmol) was dissolved in 5mL of DMSO, to which was slowly added 5mL of 3, 4-diaminophenol at room temperature with stirring(1.24g,10.0mmol) in DMSO. After 2h reaction at room temperature, a large amount of precipitate precipitated out, which was filtered off with suction and washed with ethanol to give 37(1.04g, 39.0%) as a yellow solid.¹H NMR(400MHz,DMSO-d₆)10.69(s,1H),9.44(s,1H),8.58(d,J＝9.0Hz,2H),8.42(d,J＝9.0Hz,2H),8.01(d,J＝9.1Hz,1H),7.47(dd,J＝9.1,2.7Hz,1H),7.36(d,J＝2.6Hz,1H).

Example 65: synthesis of 3- (3-nitrophenyl) quinoxalin-6-ol (38)

Following the procedure for preparation of compound 37, 2-bromo-3' -nitroacetophenone and 3, 4-diaminophenol were reacted to give 38 as a yellow solid (2.84g, 61.5%).¹H NMR(400MHz,DMSO-d₆)10.67(s,1H),9.46(s,1H),9.08–9.06(m,1H),8.78–8.74(m,1H),8.40(ddd,J＝8.2,2.3,0.9Hz,1H),8.00(d,J＝9.1Hz,1H),7.92–7.86(m,1H),7.45(dd,J＝9.1,2.7Hz,1H),7.37(d,J＝2.6Hz,1H).

Example 66: synthesis of 3- (2-nitrophenyl) quinoxalin-6-ol (39)

Following the procedure for preparation of compound 37, 2-bromo-2' -nitroacetophenone and 3, 4-diaminophenol were reacted to give 39(2.74g, 50.1%) as a yellow solid.

Example 67: synthesis of 2- (4-nitrophenyl) quinoxalin-5-ol (40)

Following the procedure for preparation of compound 37, 2-bromo-4' -nitroacetophenone and 2, 3-diaminophenol were reacted to give 40(5.93g, 44.4%) as a yellow solid.¹H NMR(400MHz,DMSO-d₆)10.56(s,1H),9.59(s,1H),8.62(d,J＝8.9Hz,2H),8.44(d,J＝8.9Hz,2H),7.78–7.72(m,1H),7.65–7.59(m,1H),7.22(dd,J＝7.7,1.0Hz,1H).

Example 68: synthesis of 2- (3-nitrophenyl) quinoxalin-5-ol (41)

White solid 41(4.57g, 64.3%) was prepared from 2-bromo-3' -nitroacetophenone and 2, 3-diaminophenol by the method of preparation of compound 37.¹H NMR(400MHz,DMSO-d₆)10.55(s,1H),9.71(s,1H),9.36–9.34(m,1H),8.95(d,J＝7.9Hz,1H),8.39(dd,J＝8.1,1.7Hz,1H),7.93–7.87(m,1H),7.75–7.69(m,1H),7.59(dd,J＝8.4,1.0Hz,1H),7.24(dd,J＝7.7,1.0Hz,1H).

Example 69: synthesis of 2- (2-nitrophenyl) quinoxalin-5-ol (42)

Following the procedure for preparation of compound 37, 2-bromo-2' -nitroacetophenone and 2, 3-diaminophenol were reacted to give 42 as a yellow solid (3.16g, 53.5%).¹H NMR(400MHz,DMSO-d₆)10.54(s,1H),9.13(s,1H),8.16(d,J＝8.1Hz,1H),7.97(d,J＝7.6Hz,1H),7.94–7.88(m,1H),7.84–7.77(m,1H),7.73–7.67(m,1H),7.45(d,J＝8.4Hz,1H),7.20(d,J＝7.7Hz,1H).

Example 70: synthesis of 7- (2-fluoroethoxy) -2- (4-nitrophenyl) quinoxaline (43)

Compound 37(388.5mg,1.5mmol), 1-bromo-2-fluoroethane (184.6mg,1.5mmol) and K2CO3 were dissolved in 5mL of anhydrous DMF and stirred at 90 ℃ under reflux for 2 h. TLC monitored the reaction for substantial completion, cooled to room temperature, added 50mL of deionized water, precipitated, filtered, washed with water, and recrystallized from methanol to yield 43(412.7mg, 90.8%) as a yellow solid.¹H NMR(400MHz,CDCl₃)9.25(s,1H),8.43(d,J＝8.9Hz,2H),8.38(d,J＝8.9Hz,2H),8.09(d,J＝9.2Hz,1H),7.54(dd,J＝9.2,2.7Hz,1H),7.47(d,J＝2.7Hz,1H),4.96–4.81(m,2H),4.49–4.39(m,2H).

Example 71: synthesis of 7- (2-fluoroethoxy) -2- (3-nitrophenyl) quinoxaline (44)

According to the procedure for the preparation of compound 43, yellow solid 44(796.6mg, 89.5%) was obtained from compound 38.¹H NMR(400MHz,CDCl₃)9.26(s,1H),9.11(s,1H),8.53(d,J＝7.8Hz,1H),8.39(dd,J＝8.1,2.1Hz,1H),8.13(d,J＝9.2Hz,1H),7.80–7.75(m,1H),7.56(dd,J＝9.2,2.7Hz,1H),7.51(d,J＝2.7Hz,1H),4.97–4.82(m,2H),4.50–4.40(m,2H).

Example 72: synthesis of 7- (2-fluoroethoxy) -2- (2-nitrophenyl) quinoxaline (45)

According to the procedure for the preparation of compound 43, compound 39 was prepared as a yellow solid 45(497.3mg, 87.7%).¹H NMR(400MHz,CDCl₃)8.85(s,1H),8.10(dd,J＝8.1,1.0Hz,1H),8.08(d,J＝9.2Hz,1H),7.81–7.73(m,2H),7.70–7.65(m,1H),7.53(dd,J＝9.2,2.8Hz,1H),7.37(d,J＝2.8Hz,1H),4.93–4.78(m,2H),4.43–4.34(m,2H).

Example 73: synthesis of 5- (2-fluoroethoxy) -2- (4-nitrophenyl) quinoxaline (46)

According to the procedure for preparation of compound 43, yellow solid 46(346.1mg, 91.3%) was obtained from compound 40.¹H NMR(400MHz,CDCl₃)9.41(s,1H),8.46–8.38(m,4H),7.84(dd,J＝8.5,1.2Hz,1H),7.80–7.75(m,1H),7.22(d,J＝7.7Hz,1H),5.06–4.91(m,2H),4.62–4.52(m,2H).

Example 74: synthesis of 5- (2-fluoroethoxy) -2- (3-nitrophenyl) quinoxaline (47)

According to the procedure for preparation of compound 43, yellow solid 47(691.2mg, 89.3%) was obtained from compound 41.¹H NMR(400MHz,CDCl₃)9.41(s,1H),9.09–9.07(m,1H),8.61(d,J＝7.8Hz,1H),8.38(dd,J＝8.2,1.3Hz,1H),7.84(d,J＝8.5Hz,1H),7.78(d,J＝8.3Hz,1H),7.74(d,J＝8.6Hz,1H),7.26–7.23(m,1H),5.06–4.92(m,2H),4.64–4.54(m,2H).

Example 75: synthesis of 5- (2-fluoroethoxy) -2- (2-nitrophenyl) quinoxaline (48)

According to the method for preparing compound 43, yellow solid 48(1.20g, 95.4%) was obtained from compound 42.¹H NMR(400MHz,CDCl₃)8.99(s,1H),8.12(dd,J＝8.1,1.0Hz,1H),7.82–7.65(m,5H),7.23–7.18(m,1H),5.05–4.90(m,2H),4.61–4.52(m,2H).

Example 76: synthesis of 4- (7- (2-fluoroethoxy) quinoxalin-2-yl) aniline (49)

Compound 43(268.0mg,0.9mmol) was dissolved in 50mL of anhydrous methanol, Pd/C catalyst (9.1mg,0.09mmol) and hydrazine hydrate (430.5mg,8.6mmol) were added, and stirring was performed at 80 ℃ for 0.5h under reflux. TLC monitored the reaction for substantial completion, the catalyst was removed by suction filtration, the solvent was removed under reduced pressure and methanol was recrystallized to yield 49 as a white solid (236.7mg, 97.2%).¹H NMR(400MHz,CDCl₃)9.12(s,1H),8.03(d,J＝8.6Hz,2H),7.98–7.94(m,1H),7.41–7.35(m,2H),6.83(d,J＝8.6Hz,2H),4.93–4.78(m,2H),4.45–4.34(m,2H),3.97(s,2H).

Example 77: synthesis of 3- (7- (2-fluoroethoxy) quinoxalin-2-yl) aniline (50)

According to the procedure for the preparation of compound 49, compound 44 was prepared as a yellow solid 50(376.2mg, 96.9%).¹H NMR(400MHz,CDCl₃)9.15(s,1H),8.01(d,J＝9.0Hz,1H),7.53–7.48(m,2H),7.45(dd,J＝9.0,2.8Hz,1H),7.42(d,J＝2.6Hz,1H),7.37–7.32(m,1H),6.87–6.82(m,1H),4.94–4.79(m,2H),4.46–4.35(m,2H),3.87(s,2H).

Example 78: synthesis of 2- (7- (2-fluoroethoxy) quinoxalin-2-yl) aniline (51)

According to the procedure for the preparation of compound 49, 51(257.9mg, 92.9%) was obtained as a yellow solid from compound 45.¹H NMR(400MHz,CDCl₃)9.17(s,1H),7.99(d,J＝9.1Hz,1H),7.80(dd,J＝8.0,1.4Hz,1H),7.42(dd,J＝9.1,2.8Hz,1H),7.32(d,J＝2.7Hz,1H),7.29–7.24(m,1H),6.90–6.80(m,2H),4.94–4.79(m,2H),4.46–4.35(m,2H).

Example 79: synthesis of 4- (5- (2-fluoroethoxy) quinoxalin-2-yl) aniline (52)

According to the method for preparing compound 49, compound 46 was prepared as a yellow solid 52(249.7mg, 92.8%).¹H NMR(400MHz,CDCl₃)9.27(s,1H),8.06(d,J＝8.7Hz,2H),7.72(dd,J＝8.5,1.1Hz,1H),7.67–7.61(m,1H),7.07(d,J＝7.8Hz,1H),6.83(d,J＝8.7Hz,2H),5.04–4.89(m,2H),4.58–4.48(m,2H),4.00(s,2H).

Example 80: synthesis of 3- (5- (2-fluoroethoxy) quinoxalin-2-yl) aniline (53)

According to the procedure for the preparation of compound 49, compound 47 gave 53 as a yellow solid (417.6mg, 92.7%).¹H NMR(400MHz,CDCl₃)9.31(s,1H),7.76(d,J＝8.4Hz,1H),7.68–7.59(m,2H),7.56(d,J＝7.8Hz,1H),7.36–7.31(m,1H),7.20(d,J＝7.8Hz,1H),6.83(dd,J＝7.9,2.2Hz,1H),5.04–4.89(m,2H),4.63–4.52(m,2H),3.90(s,2H).

Example 81: synthesis of 2- (5- (2-fluoroethoxy) quinoxalin-2-yl) aniline (54)

According to the procedure for preparation of compound 49, yellow solid 54(119.5mg, 89.7%) was obtained from compound 48.¹H NMR(400MHz,CDCl₃)9.33(s,1H),7.83(dd,J＝8.0,1.4Hz,1H),7.69–7.64(m,2H),7.28–7.24(m,1H),7.14–7.09(m,1H),6.89–6.81(m,2H),6.19(s,2H),5.04–4.90(m,2H),4.60–4.50(m,2H).

Example 82: synthesis of 4- (7- (2-fluoroethoxy) quinoxalin-2-yl) -N-methyllaniline (55) and 4- (7- (2-fluoroethoxy) quinoxalin-2-yl) -N, N-dimethyllaniline (61)

Compound 49(129.7mg,0.5mmol) was dissolved in 20mL acetone and K was added₂CO₃(127.2,0.9mmol) and methyl iodide (97.5mg,0.7mmol), the reaction was stirred at 40 ℃ overnight. TLC monitored the reaction for substantial completion, the solvent was removed under reduced pressure and the residue was chromatographed on silica gel (petroleum ether/ethyl acetate 3/1) to give yellow solids 55(31.2mg, 22.8%) and 61(45.7mg, 31.9%). 55:¹H NMR(400MHz,CDCl₃)9.14(s,1H),8.10(d,J＝8.7Hz,2H),7.96(d,J＝9.1Hz,1H),7.48(s,1H),7.37(dd,J＝9.1,2.8Hz,1H),6.76(d,J＝8.8Hz,2H),4.94–4.78(m,2H),4.46–4.35(m,2H),2.94(s,3H).¹³C NMR(101MHz,CDCl₃)159.56,152.00,151.05,143.74,140.74,137.11,130.15,128.76,125.26,121.54,112.50,107.42,82.47,81.62(d,J＝171.1Hz),67.42(d,J＝20.6Hz),30.37.61:¹H NMR(400MHz,CDCl₃)9.16(s,1H),8.17(d,J＝8.8Hz,2H),7.96(d,J＝9.1Hz,1H),7.55(s,1H),7.37(dd,J＝9.1,2.7Hz,1H),6.88(d,J＝8.5Hz,2H),4.93–4.78(m,2H),4.47–4.36(m,2H),3.09(s,63H).¹³C NMR(101MHz,CDCl₃)159.57,151.98,151.77,143.78,140.82,137.10,130.17,128.55,124.14,121.47,112.28,107.42,81.64(d,J＝171.2Hz),67.44(d,J＝20.6Hz),40.23.

example 83: synthesis of 3- (7- (2-fluoroethoxy) quinoxalin-2-yl) -N-methyllaniline (56) and 3- (7- (2-fluoroethoxy) quinoxalin-2-yl) -N, N-dimethyllaniline (62)

According to the method for preparing compounds 55 and 61, yellow solids 56(45.6mg, 36.6%) and 62(31.2mg, 23.9%) were obtained from compound 5. 56:¹H NMR(400MHz,CDCl₃)9.16(s,1H),8.01(d,J＝9.9Hz,1H),7.50–7.42(m,4H),7.41–7.35(m,1H),6.85–6.80(m,1H),4.94–4.79(m,2H),4.46–4.36(m,2H),2.97(s,3H).¹³C NMR(101MHz,CDCl₃)159.64,152.57,149.96,143.71,141.40,137.92,137.84,130.26,129.91,122.69,116.56,114.34,111.09,107.68,81.59(d,J＝171.3Hz),67.49(d,J＝20.6Hz),30.80.62:¹H NMR(400MHz,CDCl₃)9.18(s,1H),8.01(d,J＝9.9Hz,1H),7.71–7.40(m,5H),7.00(s,1H),4.94–4.79(m,2H),4.47–4.37(m,2H),3.10(s,6H).¹³C NMR(101MHz,CDCl₃)159.61,152.89,151.15,143.73,141.51,137.89,137.67,130.25,129.76,122.64,115.85,114.37,111.30,107.74,81.59(d,J＝171.3Hz),67.48(d,J＝20.6Hz),40.66.

example 84: synthesis of 2- (7- (2-fluoroethoxy) quinoxalin-2-yl) -N-methyllaniline (57)

According to the procedure for the preparation of compound 55, yellow solid 57(42.7mg, 53.2%) was obtained from compound 51.¹H NMR(400MHz,CDCl₃)9.20(s,1H),7.99(d,J＝9.1Hz,1H),7.87(dd,J＝7.9,1.2Hz,1H),7.45–7.39(m,2H),7.32(d,J＝2.7Hz,1H),6.96(d,J＝8.0Hz,1H),6.92–6.86(m,1H),4.95–4.80(m,2H),4.47–4.38(m,2H),3.02(s,3H).¹³C NMR(101MHz,CDCl₃)159.76,153.84,149.53,142.66,141.57,136.28,131.78,130.25,129.49,121.87,117.76,115.99,111.52,107.16,81.59(d,J＝171.4Hz),67.56(d,J＝20.6Hz),30.07.

Example 85: synthesis of 4- (5- (2-fluoroethoxy) quinoxalin-2-yl) -N-methyllaniline (58) and 4- (5- (2-fluoroethoxy) quinoxalin-2-yl) -N, N-dimethyllaniline (63)

Following the procedure for the preparation of compounds 55 and 61, yellow solids 58(193.7mg, 35.2%) and 63(211.4mg, 36.7%) were obtained from compound 52. 58:¹H NMR(400MHz,CDCl₃)9.28(s,1H),8.11(d,J＝8.8Hz,2H),7.73(d,J＝8.1Hz,1H),7.66–7.60(m,1H),7.06(d,J＝7.2Hz,1H),6.78(d,J＝8.7Hz,2H),5.03–4.88(m,2H),4.58–4.48(m,2H),2.94(s,3H).¹³C NMR(101MHz,CDCl₃)154.15,152.14,151.14,143.54,141.69,133.03,129.74,128.84,125.08,121.88,112.49,108.55,81.75(d,J＝171.1Hz),68.32(d,J＝21.4Hz),30.36.63:¹H NMR(400MHz,CDCl₃)9.31(s,1H),8.19(d,J＝8.8Hz,2H),7.80(d,J＝8.1Hz,1H),7.68–7.62(m,1H),7.07(d,J＝7.9Hz,1H),7.01–6.90(s,2H),5.03–4.89(m,2H),4.58–4.49(m,2H),3.10(s,6H).¹³C NMR(101MHz,CDCl₃)154.17,152.11,151.82,143.62,141.72,133.01,129.70,128.59,123.96,121.89,112.22,108.51,81.76(d,J＝171.2Hz),68.33(d,J＝21.4Hz),40.18.

example 86: synthesis of 3- (5- (2-fluoroethoxy) quinoxalin-2-yl) -N-methyllaniline (59) and 3- (5- (2-fluoroethoxy) quinoxalin-2-yl) -N, N-dimethyllaniline (64)

According to the method for preparing the compounds 55 and 61, 59(104.7mg, 32.0%) and 64(159.6mg, 46.6%) as yellow solids were obtained from the compound 53. 59:¹H NMR(400MHz,CDCl₃)9.32(s,1H),7.76(dd,J＝8.5,1.0Hz,1H),7.67–7.61(m,1H),7.61–7.58(m,1H),7.56(d,J＝7.7Hz,1H),7.41–7.35(m,1H),7.20(dd,J＝7.8,0.8Hz,1H),6.84(dd,J＝8.0,1.9Hz,1H),5.04–4.89(m,2H),4.63–4.53(m,2H),2.97(s,3H).¹³C NMR(101MHz,CDCl₃)154.40,150.93,150.02,143.74,142.56,137.59,134.88,129.83,129.16,122.15,116.45,114.23,111.79,111.30,82.04(d,J＝170.8Hz),69.27(d,J＝21.0Hz),30.70.64:¹H NMR(400MHz,CDCl₃)9.34(s,1H),7.77(d,J＝7.7Hz,1H),7.72–7.50(m,3H),7.48–7.40(m,1H),7.21(d,J＝7.5Hz,1H),7.00(s,1H),5.04–4.87(m,2H),4.64–4.52(m,2H),3.10(s,6H).¹³C NMR(101MHz,CDCl₃)154.44,151.20,151.10,143.83,142.59,137.50,134.97,129.72,129.15,122.27,115.98,114.54,112.12,111.53,82.07(d,J＝170.8Hz),69.41(d,J＝21.1Hz),40.68.

example 87: synthesis of 2- (5- (2-fluoroethoxy) quinoxalin-2-yl) -N-methyllaniline (60)

According to the procedure for the preparation of compound 55, yellow solid 60(148.4mg, 45.8%) was obtained from compound 54.¹H NMR(400MHz,CDCl₃)8.54(d,J＝7.7Hz,1H),8.02(d,J＝8.4Hz,1H),7.78–7.72(m,1H),7.63–7.57(m,1H),7.50(d,J＝8.2Hz,1H),7.45–7.39(m,1H),7.27–7.19(m,2H),5.10–4.93(m,2H),4.65–4.54(m,2H),4.04(s,3H).¹³C NMR(101MHz,CDCl₃)154.16,153.94,150.04,143.50,141.18,132.04,131.95,129.98,129.42,121.08,117.02,115.59,111.27,109.10,81.72(d,J＝171.3Hz),68.40(d,J＝21.3Hz),29.83.

A schematic of the synthesis of the compounds of examples 64-87 above is shown in FIG. 4.

Example 88:¹⁸preparation of F-labelled ligand

Firstly, an experiment step:

1) the compound (S2) - ()¹⁸F]5、(R)-[¹⁸F]5、(S)-[¹⁸F]16、(R)-[¹⁸F]16、(S)-[¹⁸F]23、(R)-[¹⁸F]23、(R,S)-[¹⁸F]30、(S,R)-[¹⁸F]30、(R,S)-[1⁸F]33 and (S, R) - [ alpha ], [¹⁸F]33 preparation of

[¹⁸F]F^-The ions were concentrated on a QMA column and eluted with 1.0mL of eluent (containing Kryptofix-2.2.213 mg,1.1mg K)₂CO₃Acetonitrile/water 4/1 general-¹⁸F]F^-Eluted from the QMA column. Adding 20mCi of fluoride ion solution into 10mL glass reaction tube, heating in 120 deg.C metal bath, and continuously introducing N₂Blowing, and adding 1.0mL of anhydrous acetonitrile for three times for azeotropic blowing to ensure that the reaction system is anhydrous. 3.0mg of the labeled precursor [ Compounds S-12, R-12, S-20, R-20, S-27, R-27, (R, S) -29, (S, R) -29, (R, S) -32 and (S, R) -32, respectively ]]Dissolved in 600. mu.L of anhydrous acetonitrile, and the solution is transferred to a solution containing [ 2 ], [¹⁸F]F^-In the glass reaction tube. The reaction was heated at 100 ℃ for 6 minutes. After cooling, 400. mu.L of hydrochloric acid solution (1M) was added and the reaction was continued at 100 ℃ for 5 minutes to remove the Boc protecting group. After cooling, saturated NaHCO was added₃The reaction system was made slightly alkaline with solution and 10mL of deionized water was added to dilute the reaction mixture. The mixed solution is purified by a pretreated Sep-Pak Plus PS-2 solid phase extraction cartridge, and the cartridge is rinsed with 20mL of deionized water to remove unreacted [ alpha ], [ beta ] -n¹⁸F]F-and inorganic salts. Eluting the column with anhydrous acetonitrile (washing 3 times, each time 1mL), eluting the labeled compound and labeled precursor adsorbed on the column, concentrating, and separating and purifying by HPLC, wherein the separation conditions are as follows: venusil MP C18 reverse column (5 μm,10 mm. times.250 mm), collecting the effluent of the target product, removing the solvent by rotary evaporation, dissolving the obtained product in 10% ethanol and formulating with purified water to the desired concentration.

II, experimental results:

(S)-[¹⁸F]5、(R)-[¹⁸F]5、(S)-[¹⁸F]16、(R)-[¹⁸F]16、(S)-[¹⁸F]23、(R)-[¹⁸F]23、(R,S)-[¹⁸F]30、(S,R)-[¹⁸F]30、(R,S)-[1⁸F]33 and (S, R) - [ alpha ], [¹⁸F]The labeling rate of 33 was 20-30%. After separation and purification by HPLC, radiochemistryAll greater than 95% pure and consistent with the retention time of the stable fluoro ligands (table 1).

TABLE 1¹⁸Retention time and purity of F-labeled ligands and their stabilizing ligands

Experimental example 89: fluorescent staining experiment

Firstly, an experiment step:

fluorescent staining of brain sections of AD patients

(1) Respectively preparing aqueous solution (containing 20% ethanol) of the compound with the concentration of 1 μ M;

(2) two AD human brain slices (paraffin) with the thickness of 8 mu m are sequentially subjected to xylene dewaxing for 15min, then sequentially subjected to 100% ethanol for 2 x 5min, 95% ethanol for 2 x 5min, 80% ethanol for 5min and 70% ethanol for 5min, washed by running water for 10min, and then placed in 10mM PBS (pH 7.4);

(3) soaking brain slices in the compound solution for 15 min;

(4) the sections were washed with 40% ethanol, rapidly differentiated and observed using a fluorescence microscope.

II, experimental results:

the experimental result is shown in fig. 5, the probe of the invention can clearly mark the neurofibrillary tangle NFTs on the slice of the AD human brain, but does not bind to the Abeta plaque or has weak binding, which indicates that the probe provided by the invention selectively binds to Tau protein.

Table 2 staining evaluation and quantitative activity data of fluoro compounds on AD human brain sections with tau and Α β

Experimental example 90: autoradiography experiment

After the labeled products with certain concentration are combined with plaques in brain slices of AD patients respectively, the plaques are exposed through a phosphorus screen, and then the images are analyzed by a phosphorus screen storage system.

Firstly, an experiment step:

(1) preprocessing an AD human brain slice;

(2) covering 20 μ Ci on AD human brain sections¹⁸F labeled compound solution 100 u L, room temperature incubation for 40 minutes;

(3) washing with 20% ethanol solution for 1 min;

(4) after drying, the preservative film is coated and placed under a phosphorus screen for exposure for 40min, and the image is analyzed by a phosphorus screen storage system.

II, experimental results:

the experimental result is shown in fig. 6, which fully indicates that the compound of the invention can be used as an AD brain tau protein imaging agent after being labeled by radionuclide, and has potential application prospect in clinical diagnosis.

Experimental example 91: in vivo biodistribution experiment in Normal mice

Pharmacokinetic properties in mice, in particular initial brain uptake and brain clearance, were studied by in vivo profiling experiments.

Firstly, an experiment step:

a labeled compound (5-10 μ Ci) (100 μ L of physiological saline solution containing 10% ethanol) was injected into normal mice (ICR, male, 20-22g, 5 weeks old) from the tail vein (n ═ 5), decapitated at 2 minutes, 10 minutes, 30 minutes, and 60 minutes after injection, respectively, and relevant organs were dissected out and measured for wet weight and radioactive counts. Data are expressed as percent radioactivity (% ID/g) per gram of organ.

II, experimental results:

as shown in Table 3, the probe (S) -, [ 2 ] of the present invention¹⁸F]5、(R)-[¹⁸F]5、(S)-[¹⁸F]16、(R)-[¹⁸F]16、(S)-[¹⁸F]23、(R)-[¹⁸F]23、(R,S)-[¹⁸F]30、(S,R)-[¹⁸F]30、(R,S)-[1⁸F]33 and (S, R) - [ alpha ], [¹⁸F]33 can smoothly pass through the blood brain barrier, the brain uptake reaches the peak value in 2 minutes, and the brain is cleared quickly in normal mice.

Table 3:¹⁸results of biodistribution of F-labeled Compounds in Normal mice

a is expressed as% ID/g, n is 4-5

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Claims (5)

1. 2-arylquinoxaline compounds having affinity with Tau protein, characterized in that the compounds are any of compounds 4) to 7):

wherein F in the formulae 4) to 7) is¹⁸F or¹⁹F。

2. A derivative of the compound of claim 1 which is a pharmaceutically acceptable salt.

3. A diagnostic or diagnostic reagent for a neurofibrillary tangle disease caused by Tau protein deposition, characterized in that the effective ingredient is the compound according to claim 1 and/or the derivative according to claim 2.

4. The diagnostic or detection reagent of claim 3, wherein the disease comprises Alzheimer's disease, frontotemporal lobe degeneration, chronic traumatic encephalopathy, progressive supranuclear palsy, corticobasal degeneration or pick's disease.

5. Use of a compound according to claim 1 or a derivative according to claim 2 for the preparation of a nuclear medicine imaging agent or an optical imaging agent.

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