Chiral spirocyclic pentanoperisoxazoline compounds and uses thereof

Abstract

The application discloses a kind of chiral spiro cyclopentane and isoxazoline compound in the technical field of organic synthesis chemistry and medicinal chemistry, its structure general formula is as follows: Wherein, the spiro ring skeleton can independently be represented as benzopentazacyclic ring, benzopentacarbon ring, benzopentaoxygen heterocycle or benzohexazacyclic ring.Studies show that chiral spiro cyclopentane and isoxazoline compound can effectively inhibit lipopolysaccharide-induced macrophage RAW264.7 release inflammatory factor NO, provide its application in preparing anti-inflammatory drugs.

Description

Technical Field

[0001] This invention relates to the fields of organic synthetic chemistry and medicinal chemistry, specifically to chiral spirocyclopentane-isooxazoline compounds and their applications. Background Technology

[0002] Isoxazoline compounds are a class of five-membered heterocyclic pharmacological skeletons with both nitrogen and oxygen heteroatoms. They possess strong chemical modifiability and excellent biological activity, making them one of the core heterocyclic structures in organic synthesis and drug development. They can be derived into various cyclic and acyclic compounds and are also key precursors for the synthesis of amino alcohols, hydroxy ketones, isoxazolidines, and other similar compounds. Studies have found that compounds containing isoxazoline pharmacological groups typically exhibit significant activity and are highly favored in drug development. Due to their anti-inflammatory and antibacterial pharmacological activities, they play an important role in the synthesis of natural products and the preparation of various pharmacologically active heterocyclic compounds, attracting widespread attention in the scientific community.

[0003] According to literature reports, isoxazoline hydrazides can effectively inhibit histamine H1 receptors, blocking allergic inflammation caused by them; a series of isoxazoline derivatives of thiophene and furan compounds have a strong inhibitory effect on 5-lipoxygenase, thereby inhibiting the synthesis of pro-inflammatory mediators. Methods for synthesizing five-membered cyclic isoxazolines are mainly divided into two types: metal-catalyzed and non-metal-catalyzed. Metal-free catalytic reactions include the 1,3-dipolar cycloaddition reaction of benzoylnitrosomethane / ethyl 2-nitroacetate with terminal alkenes; metal-free one-pot synthesis of isoxazolines; cycloaddition reaction of five-membered cyclic nitrosones with DMAD; transition metal-catalyzed reactions, such as the addition / cyclization tandem reaction of N-methyl-tert-butyl-substituted cyclic nitrones with alkynes; the 1,3-dipolar cycloaddition reaction of aldoximes with alkenes; and the 1,3-dipolar cycloaddition reaction of N-methyl nitrones with electron-deficient alkenes. These studies provide effective pathways for the synthesis of isoxazoline derivatives.

[0004] Despite numerous reports on the synthesis of isoxazoline compounds, research on the synthesis of these chiral molecules is still scarce. Currently, there is only one reported case of constructing chiral isoxazoline-cyclopentane compounds via an organocatalyzed asymmetric dearomatization cyclization reaction of allenoic acid esters and nitroisoxazole. Chiral spirocyclopentane-isoxazoline derivatives obtained by transition metal-catalyzed asymmetric dearomatization of nitroisoxazole, representing a novel skeletal structure, have not been publicly reported, and no anti-inflammatory activity evaluations have been conducted on these compounds. Currently, inflammation-related diseases and bacterial infections remain common clinical problems, and existing anti-inflammatory and antibacterial drugs still suffer from shortcomings such as drug resistance, poor selectivity, and significant side effects. Developing chiral isoxazoline drug candidates with novel skeletal structures and exploring their anti-inflammatory and antibacterial pharmacological effects is of significant practical importance for enriching the drug candidate library. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention designs several novel chiral spirocyclopentane-isooxaoxolin structures.

[0006] One objective of this invention is to provide several novel chiral spirocyclopentane-isooxaline compounds, with the following general structural formula: , In the formula: the spirocyclic skeleton can be independently represented as a benzo5-membered nitrogen heterocycle, a benzo5-membered carbon ring, a benzo5-membered oxygen heterocycle, or a benzo6-membered nitrogen heterocycle.

[0007] Furthermore, (1) when the spirocyclic skeleton is a benzo5-membered nitrogen heterocycle, the general formula of the compound is as follows: , Ar 1 It is independently selected from phenyl, 2-fluorophenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-ethylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-trifluoromethylphenyl, 4-methoxycarbonylphenyl, 4-phenylphenyl or 3-chloro-4-methoxyphenyl; Accordingly, Ar 2 It is independently selected from phenyl, 4-chlorophenyl, 4-bromophenyl, 5-methylphenyl, 5-methoxyphenyl, 5-chlorophenyl, 5-bromophenyl, 6-chlorophenyl, 6-bromophenyl, 7-methylphenyl, 7-chlorophenyl, 7-bromophenyl, 4-bromo-5-methylphenyl, 5,7-dimethylphenyl or 5-bromo-7-methylphenyl; Accordingly, R is independently selected from methyl, benzyl, or phenyl; (2) When the spirocyclic skeleton is a benzo5-membered carbon ring, the general formula of the compound is as follows: ; (3) When the spirocyclic skeleton is a benzo5-membered oxygen heterocycle, the general formula of the compound is as follows: ; (4) When the spirocyclic skeleton is a benzo-6-membered nitrogen heterocycle, the general formula of the compound is as follows: .

[0008] Furthermore, the structural formula of the compound is as follows: , .

[0009] The second objective of this invention is to provide its application in the preparation of anti-inflammatory drugs based on the fact that chiral spirocyclopentane-isooxazoline compounds can effectively inhibit the release of inflammatory factor NO from lipopolysaccharide-induced macrophages RAW264.7. Attached Figure Description

[0010] Figure 1 The chiral compound 3a prepared in Example 20 1 H NMR spectrum; Figure 2 The chiral compound 3a prepared in Example 20 13 C NMR spectrum; Figure 3 The high-performance liquid chromatogram of exotropic compound 3a prepared in Example 20; Figure 4 The high-performance liquid chromatogram of chiral compound 3a prepared in Example 20; Figure 5 The chiral compound 3c prepared in Example 22 1 H NMR spectrum; Figure 6 The chiral compound 3c prepared in Example 22 13 C NMR spectrum; Figure 7 The high-performance liquid chromatogram of the exotropic compound 3c prepared in Example 22; Figure 8 The high-performance liquid chromatogram of chiral compound 3c prepared in Example 22; Figure 9 The single-crystal diffraction pattern of chiral compound 3j prepared in Example 29; Figure 10 The single-crystal diffraction pattern of chiral compound 5 prepared in Example 54; Figure 11 The chiral compound 9 prepared in Example 56 1 H NMR spectrum; Figure 12 The chiral compound 9 prepared in Example 56 13 C NMR spectrum; Figure 13 The high-performance liquid chromatogram of exotropic compound 9 prepared in Example 56; Figure 14 The high-performance liquid chromatogram of chiral compound 9 prepared in Example 56. Detailed Implementation

[0011] The method for preparing the novel chiral spirocyclopentane-isoxazolino structure involved in this invention includes the following steps: Step 1: Add 3-aryl-4-nitroisoxazole and rhodium acetate dimer sequentially to the reaction tube, then add an organic solvent to obtain a mixture. Heat the mixture at 25°C. o Stirred at C for 1 h, then add palladium catalyst, chiral phosphine ligand, and vinylcyclopropanes with different ring substitutions in sequence. The mixture is then stirred at 40 °C.o C~-10 o C was stirred under reaction temperature and inert gas protection; Step 2: After the reaction is complete, the corresponding chiral spirocyclopentane and isoxazoline compounds are obtained through post-processing and separation.

[0012] For example: First, add 0.20 mmol of 3-aryl-4-nitroisoxazole and 0.002 mmol of Rh2(OAc)4 to 2.5 mL of dichloromethane, and then mix the mixture at 25°C. o The mixture was stirred at C for 1 h, and then 0.01 mmol of Pd2(dba)3, 0.02 mmol of L4, L6, or L8 ligands, and 0.44 mmol of vinylcyclopropane with different ring substitutions were added sequentially to the mixture. The mixture was then incubated at -10 °C. o The mixture was stirred for 48 h under the protection of C and argon. After the reaction was completed, the mixture was directly separated by column chromatography to obtain the corresponding chiral spirocyclopentane-isooxazoline compounds.

[0013] The method for preparing novel structures of chiral spirocyclopentane-isooxazoline compounds of the present invention utilizes 3-aryl-4-nitroisooxazoles with different structures and vinylcyclopropanes with different structures to synthesize different types of chiral spirocyclopentane-isooxazoline 3a~z, 3a'~h', 5~9, with specific structural formulas as follows: , .

[0014] In this invention, the yield of the product is the separation yield, and the structure of the product is confirmed by nuclear magnetic resonance and X-ray single crystal diffraction.

[0015] Examples 1-20:

[0016] In a 10 mL dry reaction tube, add 0.20 mmol of 3-phenyl-4-nitroisoxazole 1a, 0.002 mmol of Rh2(OAc)4 (or without Rh2(OAc)4), 0.01 mmol of palladium catalyst, 0.02 mmol of chiral phosphine ligand, and 0.24 mmol of spiro[vinylcyclopropane-] N [-substituted indole oxide]2 was added to 2 mL of organic solvent, and the mixture was stirred under different temperatures and argon protection. The reaction progress was monitored by thin-layer chromatography. After the reaction was completed, the mixture was purified by column chromatography to obtain product 3 (the results of the examples are shown in Table 1 below).

[0017]

[0018] Note: a: Separation yield; b: Confirmed by high performance liquid chromatography analysis; c: 0.1 mmol1a was used; d: 0.1 mmol1a and 0.22 mmol2 were used. DCM: dichloromethane; toluene: toluene; THF: tetrahydrofuran; MTBE: methyl tert-butyl ether.

[0019] Examples 20-53: ; In a 10 mL dry reaction tube, 0.20 mmol of 3-aryl-4-nitroisoxazole 1 and 0.002 mmol of Rh2(OAc)4 were first added to 2.5 mL of dichloromethane. The mixture was then incubated at 25 °C. o The mixture was stirred at C for 1 h, and then 0.01 mmol of Pd2(dba)3, 0.02 mmol of L4, L6, or L8 ligands, and 0.44 mmol of vinylcyclopropane with different ring substitutions were added sequentially to the mixture. The mixture was then incubated at -10 °C. o The mixture was stirred for 48 h under C and argon protection, and the reaction progress was monitored by thin-layer chromatography. After the reaction was completed, the mixture was purified by column chromatography to obtain product 3 (the results of the examples are shown in Table 2 below).

[0020]

[0021] Note: a: Separation yield; b: Confirmed by high performance liquid chromatography analysis.

[0022] The characterization data of products 3a~z and 3a'~h' in the examples are as follows: Characterization data of product 3a: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 5 / 95 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o C); majordiastereomer: t R (major) = 21.4 min; 1 H NMR (400 MHz, CDCl3): δ 7.87 (d, J = 7.2 Hz,2H), 7.53-7.46 (m, 3H), 7.36-7.31 (m, 1H), 6.97 (t, J = 7.6 Hz, 1H), 6.90 (d,J =8.0 Hz, 1H), 6.82 (d, J = 7.2 Hz, 1H), 6.17 (s, 1H), 6.00-5.91 (m, 1H), 5.42(d, J = 10.0 Hz, 1H), 5.33 (d, J = 16.8 Hz, 1H), 3.72-3.66 (m, 1H), 3.26 (s, 3H),2.85 (t, J = 12.8 Hz, 1H), 2.08 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.8, 153.4, 143.8, 133.0, 131.1, 129.5, 128.9, 128.8, 128.5, 128.4 127.0,126.4, 124.0, 122.7, 122.6, 110.1, 108.9, 98.0, 58.9, 50.6, 43.3, 26.8. HRMS(ESI-TOF) m / z : [M + H] + calcd for C 22 H 20 N3O4390.1454; found: 390.1453. Characterization data of product 3b: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o C); major diastereomer: t R (major) = 41.7 min; 1 H NMR (400 MHz, CDCl3): δ 7.87 (d, J = 6.8 Hz,2H), 7.52-7.46 (m, 3H), 7.31-7.22 (m, 6H), 6.93 (t, J = 7.2 Hz, 1H), 6.80 (dd, J= 18.4, 7.2 Hz, 2H), 6.27 (s,1H), 6.02-5.93 (m, 1H), 5.43 (d, J = 10.0 Hz, 1H),5.33 (d, J = 16.8 Hz, 1H), 4.97 (s, 2H), 3.75-3.68 (m, 1H), 2.94 (t, J = 12.8 Hz,1H), 2.15 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 176.1, 153.4, 142.9,135.2, 133.1, 131.1, 129.4, 129.0, 128.9, 129.9, p128.6, 128.5, 127.8 127.4,127.1, 127.1, 126.4, 124.1, 122.8, 122.7, 110.1, 110.0, 98.2, 59.0, 50.8,44.3, 43.4. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 28 H 24 N3O4466.1767; found:466.1771. Characterization data of product 3c: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o C); major diastereomer: t R (minor) = 23.2 min, t R (major) = 35.6 min; 1 H NMR (400 MHz, CDCl3): δ 7.89 (d, J= 7.2 Hz, 2H), 7.55-7.47 (m, 5H), 7.44-7.42 (m, 3H), 7.24-7.22 (m, 1H), 7.00 (t, J = 7.6 Hz, 1H), 6.87 (dd, J = 7.2, 4.0 Hz, 2H), 6.26 (s,1H), 6.03-5.94 (m, 1H), 5.42 (d, J = 10.4 Hz, 1H), 5.33 (d, J = 17.2 Hz, 1H),3.77-3.70 (m, 1H), 2.94 (t, J = 13.2 Hz, 1H), 2.23 (q, J = 12.8 Hz, 1H). 13 C NMR(101 MHz, CDCl3): δ 175.5, 153.5, 143.9, 134.1, 133.1, 131.2, 129.8, 129.6,129.4, 129.0, 129.0, 128.6, 127.0, 126.7, 126.5, 124.4, 123.1, 122.8, 110.2,98.3, 59.2, 50.8, 43.8. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 22 N3O4 452.1613; found: 452.1610. Characterization data of product 3d: The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (major) = 11.2 min; 1 H NMR (400 MHz, CDCl3): δ 7.85 (d, J= 6.8 Hz, 2H), 7.46-7.40 (m, 3H), 7.30 (t, J = 8.0 Hz, 1H), 7.09 (d, J = 8.0 Hz, 1H), 6.77 (d, J = 7.6 Hz, 1H), 6.52 (s, 1H), 5.90-5.82 (m, 1H), 5.34 (d, J = 10.0 Hz, 1H), 5.22 (d, J = 16.8 Hz, 1H), 4.39-4.33 (m, 1H), 3.37 (t, J = 13.2 Hz, 1H), 3.18 (s, 3H), 2.13 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 174.5, 154.0, 145.8, 133.4, 130.8, 130.7, 130.6, 129.3, 128.6, 126.9, 124.5, 123.3, 122.6, 110.9, 107.3, 94.4, 60.4, 48.7, 38.4, 26.8. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 22 H 19 ClN3O4 424.1064; found: 424.1067. Characterization data of product 3e: The ee was determined by HPLC (Chiralpak AD-3 column, i -PrOH / hexane = 4 / 96 v / v, flow rate 0.8 mL / min, λ = 214 nm, 25 o °C); major diastereomer: t R (major) = 39.0 min; 1 H NMR (400 MHz, CDCl3): δ 7.84 (d, J= 7.2 Hz, 2H), 7.46-7.41 (m, 3H), 7.28-7.21 (m, 2H), 6.81 (d, J = 7.2 Hz, 1H), 6.60 (s, 1H), 5.93-5.84 (m, 1H), 5.35 (d, J = 10.0 Hz, 1H), 5.22 (d, J = 17.2 Hz, 1H), 4.41-4.35 (m, 1H), 3.45 (t, J = 12.8 Hz, 1H), 3.18 (s, 3H), 2.12 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 174.6, 154.1, 146.1, 133.5, 131.1, 130.7, 129.3, 128.6, 127.7, 126.9, 124.6, 122.6, 118.9, 110.9, 107.8, 94.1, 61.1, 48.5, 38.4, 26.7. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 22 H 19 BrN3O4 468.0559; found: 468.0561. Characterization data of product 3f: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 3 / 97 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o C); major diastereomer: t R (major) = 24.5 min; 1 H NMR (400 MHz, CDCl3): δ 7.87 (d, J = 6.8 Hz, 2H), 7.56-7.48 (m, 3H), 7.13 (d, J = 7.6 Hz, 1H), 6.79 (d, J= 8.0 Hz, 1H), 6.59(s, 1H), 6.16 (s, 1H), 5.99-5.91 (m, 1H), 5.42 (d, J = 10.0 Hz, 1H), 5.33 (d, J =17.2 Hz, 1H), 3.67-3.60 (m, 1H), 3.25 (s, 3H), 2.82 (t, J = 13.2 Hz, 1H), 2.23(s, 3H), 2.06 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.7, 153.6,141.5, 133.2, 132.0, 131.2, 129.8, 129.1, 129.0, 127.2, 126.5, 125.2, 122.7,110.1, 108.6, 98.2, 59.0, 50.3, 43.0, 26.9, 21.2. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 23 H 22 N3O4404.1610; found: 404.1612. Characterization data of product 3g: The ee was determined by HPLC (Chiralpak OD-3 column, i -PrOH / hexane = 7 / 93 v / v, flow rate 1.0 mL / min, λ = 214 nm, 25 o C); major diastereomer: t R (major) = 16.8 min; 1 H NMR (400 MHz, CDCl3): δ 7.86 (d, J = 6.8 Hz,2H), 7.56-7.48 (m, 3H), 7.32 (dd, J = 8.4, 1.6 Hz, 1H), 6.80 (dd, J= 18.4, 8.0Hz, 2H), 6.14 (s, 1H), 5.99-5.90 (m, 1H), 5.42 (d, J = 10.0 Hz, 1H), 5.32 (d, J =17.2 Hz, 1H), 3.61-3.54 (m, 1H), 3.26 (s, 3H), 2.83 (t, J = 13.2 Hz, 1H), 2.07(q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.5, 153.9, 142.6, 132.9,131.4, 129.5, 129.3, 129.1, 128.8, 128.0, 126.1, 124.9, 123.0, 110.0, 109.7,98.1, 59.1, 50.4, 42.9, 27.1. HRMS (ESI-TOF) m / z : [M + H] + calcd forC 22 H 19 ClN3O4424.1064; found: 424.1073. Characterization data of product 3h:. The ee was determined by HPLC (Chiralpak AD-3 column, i -PrOH / hexane = 7 / 93 v / v, flow rate 0.8 mL / min, λ = 214 nm, 25 o C); majordiastereomer: t R (minor)= 24.3 min, t R (major) = 22.0 min; 1 H NMR (400 MHz,CDCl3): δ 7.87 (d, J = 7.2 Hz, 2H), 7.55-7.45 (m, 4H), 6.91 (s, 1H), 6.78 (d, J =8.4 Hz, 1H), 6.14 (s, 1H), 5.98-5.89 (m, 1H), 5.42 (d,J = 10.0 Hz, 1H), 5.32(d, J = 17.2 Hz, 1H), 3.60-3.53 (m, 1H), 3.26 (s, 3H), 2.82 (t, J = 13.2 Hz, 1H),2.07 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.4, 154.0, 143.1, 132.9,132.4, 131.5, 129.4, 129.2, 129.1, 128.6, 127.6, 126.1, 123.0, 115.3, 110.2,110.0, 98.2, 59.0, 50.3, 42.8, 27.1. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 22 H 19 BrN3O4 468.0559; found: 468.0560. Characterization data of product 3i: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (minor)= 20.6 min, t R (major) = 33.9 min; 1 H NMR (400 MHz, CDCl3): δ 7.85 (d, J = 6.4 Hz, 2H), 7.54-7.49 (m, 3H), 6.95 (d, J = 7.6 Hz, 1H),6.90 (s, 1H), 6.71 (d, J = 7.6 Hz, 1H), 6.15 (s, 1H), 6.00-5.91 (m, 1H), 5.44(d, J= 10.0 Hz, 1H), 5.33 (d, J = 17.2 Hz, 1H), 3.64-3.59 (m, 1H), 3.26 (s, 3H),2.84 (t, J = 12.8 Hz, 1H), 2.06 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.9, 153.6, 145.2, 135.5, 133.0, 131.3, 129.1, 128.9, 126.3, 125.6, 125.0,123.0, 122.5, 110.0, 109.7, 98.0, 58.7, 50.8, 43.2, 27.1. HRMS (ESI-TOF) m / z :[M + H] + calcd for C 22 H 19 ClN3O4424.1064; found: 424.1065. Characterization data of product 3j: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 3 / 97 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (minor)= 25.9 min, t R (major) = 47.8 min; 1 H NMR (400 MHz, CDCl3): δ 7.85 (d, J = 6.8 Hz, 2H), 7.55-7.46 (m, 3H), 7.10 (d, J = 8.0 Hz, 1H),7.05 (s, 1H), 6.65 (d, J = 8.0 Hz, 1H), 6.14 (s, 1H), 5.99-5.90 (m, 1H), 5.43(d, J = 10.0 Hz, 1H), 5.32 (d,J = 17.2 Hz, 1H), 3.65-3.59 (m, 1H), 3.25 (s, 3H), 2.83 (t, J = 12.8 Hz, 1H), 2.06 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.8, 153.5, 145.3, 133.0, 131.3, 129.1, 128.9, 126.3, 126.1, 125.4, 125.3, 123.3, 123.0, 112.5, 110.0, 97.9, 58.8, 50.8, 43.1, 27.1. HRMS (ESI-TOF) m / z :[M + H] + calcd for C 22 H 19 BrN3O4 468.0559; found: 468.0557. Characterization data of product 3k: The ee was determined by HPLC (Chiralpak OD-3 column, i i-PrOH / hexane = 7 / 93 v / v, flow rate 1.0 mL / min, λ = 214 nm, 25 o °C); major diastereomer: t R (major) = 10.6 min; 1 H NMR (400 MHz, CDCl3): δ 7.86 (d, J = 7.2 Hz, 2H), 7.53-7.46 (m, 3H), 7.06 (d, J = 7.6 Hz, 1H), 6.86 (t, J = 7.2 Hz, 1H), 6.64 (d, J = 7.2 Hz, 1H), 6.16 (s, 1H), 6.01-5.92 (m, 1H), 5.41 (d, J = 10.0 Hz, 1H), 5.31 (d, J= 16.8 Hz, 1H), 3.71-3.64 (m, 1H), 3.57 (s, 3H), 2.85 (t, J = 12.8 Hz,1H), 2.59 (s, 3H), 2.06 (q, J = 14.0 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 176.9,153.5, 141.7, 133.4, 131.1, 129.4,129.0, 128.6, 127.9, 126.6, 122.7, 122.5,122.1, 120.6, 110.4, 98.5, 58.4, 50.8, 43.9, 30.5, 19.4. HRMS (ESI-TOF) m / z :[M + H] + calcd for C 23 H 22 N3O4404.1610; found: 404.1613. Characterization data of product 3l: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (major) = 17.6 min; 1 H NMR (400 MHz, CDCl3): δ 7.86 (d, J = 7.2 Hz,2H), 7.54-7.47 (m, 3H), 7.27 (d, J = 7.6 Hz, 1H), 6.89 (t, J = 7.6 Hz, 1H), 6.70(d, J = 7.2 Hz, 1H), 6.16 (s, 1H), 6.01-5.92 (m, 1H), 5.44 (d, J = 10.0 Hz, 1H),5.33 (d, J= 16.8 Hz, 1H), 3.67 (s, 3H), 3.66-3.57 (m, 1H), 2.86 (t, J = 13.2 Hz,1H), 2.09 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 176.5, 153.5, 139.9,133.1, 131.9, 131.3, 130.0, 129.4, 129.0, 128.9, 128.6, 126.3, 123.4, 122.9,122.7, 116.5, 110.2, 98.3, 58.7, 50.8, 43.8,30.6. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 22 H 19 ClN3O4424.1064; found: 424.1060. Characterization data of product 3m: The ee was determined by HPLC (Chiralpak AD-3 column, i -PrOH / hexane = 13 / 87 v / v, flow rate 0.8 mL / min, λ = 214 nm, 25 o °C); major diastereomer: t R (major) = 12.8 min; 1 H NMR (400 MHz, CDCl3): δ 7.85 (d, J = 7.2 Hz,2H), 7.53-7.44 (m, 4H), 6.82 (t, J = 7.6 Hz, 1H), 6.73 (d, J = 7.2 Hz, 1H), 6.16(s, 1H), 6.00-5.92 (m, 1H), 5.43 (d, J = 10.0 Hz, 1H), 5.32 (d, J = 17.2 Hz, 1H),3.67 (s, 3H), 3.66-3.58 (m, 1H), 2.86 (t,J = 12.8 Hz, 1H), 2.09 (q, J = 13.2 Hz,1H). 13 C NMR (101 MHz, CDCl3): δ 176.7, 153.5, 141.3, 135.3, 133.1, 131.3, 130.3,129.3, 129.0, 128.9, 128.6, 126.3, 123.7, 123.2, 122.9, 110.3, 103.3, 98.3,58.6, 50.8, 43.8, 30.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 22 H 19 BrN3O4 468.0559; found: 468.0561. Characterization data of product 3n: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (major) = 15.8 min; 1 H NMR (400 MHz, CDCl3): δ 7.83 (d, J = 7.2 Hz,2H), 7.45-7.39 (m, 4H), 7.23 (d, J = 8.0 Hz, 1H), 6.71 (d, J = 7.6 Hz, 1H), 6.65(s, 1H), 5.93-5.84 (m, 1H), 5.33 (d, J = 10.0 Hz, 1H), 5.21 (d, J = 16.8 Hz, 1H),4.43-4.36 (m, 1H), 3.51 (t, J = 12.8 Hz, 1H), 3.14 (s, 3H), 2.40 (s, 3H), 2.09(q, J= 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 174.5, 154.0, 143.6, 133.6,133.1, 131.3, 130.5, 129.2, 128.5, 126.9, 124.7, 122.4, 121.6, 110.9, 107.5,93.8, 61.6, 48.3, 38.2, 26.6, 22.6. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 23 H 21 BrN3O4 482.0715; found: 482.0714. Characterization data of product 3o: The ee was determined by HPLC (Chiralpak AD-H column, i i-PrOH / hexane = 2 / 98 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (major) = 29.9 min; 1 H NMR (400 MHz, CDCl3): δ 7.86 (d, J = 7.2 Hz, 2H), 7.54-7.48 (m, 3H), 6.88 (d, J = 9.2 Hz, 1H), 6.41 (s, 1H), 6.15 (s, 1H), 5.99-5.90 (m, 1H), 5.41 (d, J = 10.0 Hz, 1H), 5.32 (d, J = 17.2 Hz, 1H), 3.67-3.61 (m, 1H), 3.54 (s, 3H), 2.82 (t, J = 12.8 Hz, 1H), 2.54 (s, 3H), 2.17 (s, 3H), 2.05 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ176.6, 153.6, 139.2, 133.8, 133.3, 131.7, 131.2, 129.1, 129.0, 128.6, 127.8, 126.5, 123.0, 122.6, 120.2, 110.2, 98.5, 58.4, 50.2, 43.4, 30.4, 29.8, 20.8, 19.2. HRMS (ESI-TOF) m / z : [M+ H] + calcd for C 24 H 24 N3O4418.1767; found: 418.1766. Characterization data of product 3p: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o C); major diastereomer: t R (major) = 24.8 min; 1 H NMR (400 MHz, CDCl3): δ 7.74 (t, J = 7.2 Hz, 2H), 7.45-7.38 (m, 3H), 7.12 (d, J = 8.4 Hz, 1H), 6.62 (s, 1H), 6.01 (s, 1H), 5.86-5.78 (m, 1H), 5.30 (d, J = 10.0 Hz, 1H), 5.20 (d, J = 17.2 Hz, 1H), 3.49-3.47 (m, 1H), 3.43 (s, 3H), 2.70 (t, J = 13.2 Hz, 1H), 2.45 (s, 3H), 1.94 (q, J =12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ176.3, 153.9, 141.0, 135.7, 132.9,131.4, 129.7, 129.4, 129.3, 129.1,128.6, 126.0, 125.3, 122.9, 122.4, 114.9,110.2, 98.5, 58.4, 50.2, 43.2, 30.5, 19.1. HRMS(ESI-TOF) m / z : [M + H] + calcd for C 23 H 21 BrN3O4 482.0715; found: 482.0718. Characterization data of product 3q: The ee was determined by HPLC (Chiralpak AD-H column, i i-PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ λ = 254 nm, 25 o °C); major diastereomer: t R (minor)= 13.3 min, t R (major) = 16.8 min; 1 1H NMR (400 MHz, CDCl3): δ 7.90 (t, J J = 7.2 Hz, 2H), 7.55 (t, J J = 8.4 Hz, 5H), 7.44 (d, J J = 6.8 Hz, 3H), 7.07 (d, J J = 8.0 Hz, 1H), 6.78 (d, J J = 8.0 Hz, 1H), 6.67 (s, 1H), 6.27 (s, 1H), 6.03-5.94 (m, 1H), 5.44 (d, J J = 10.0 Hz, 1H), 5.36 (d, J J = 17.2 Hz, 1H), 3.74-3.66 (m, 1H), 2.93 (t, J J = 12.8 Hz, 1H), 2.25 (s, 3H), 2.22-2.13 (m, 1H). 13C NMR (101 MHz, CDCl3): δ 175.4, 153.7, 141.6, 134.3, 133.2, 132.6, 131.3,129.8, 129.7, 129.1, 129.1, 128.4, 127.0, 126.6, 126.5, 125.4, 122.8, 121.0,119.3, 110.2, 110.0, 98.4, 59.2, 50.3, 43.5, 21.2. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 28 H 24 N3O4466.1767; found: 466.1773. Characterization data of product 3r: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 4 / 96 v / v, flow rate 1.0 mL / min, λ = 210 nm, 25 o C); major diastereomer: t R (major) = 53.8 min; 1 H NMR (400 MHz, CDCl3): δ 7.90 (d, J = 7.2 Hz, 2H), 7.52-7.42 (m, 8H), 6.78 (q, J = 14.8 Hz, 2H), 6.49 (s, 1H), 6.26 (s, 1H), 6.02-5.93 (m, 1H), 5.42 (d, J = 10.0 Hz, 1H), 5.34 (d, J = 17.2 Hz, 1H), 3.74-3.67 (m, 1H), 3.64 (s, 3H), 2.93 (t, J = 12.8 Hz, 1H), 2.22 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ175.1, 156.0, 153.5, 137.2, 134.3, 133.0, 131.2, 129.7, 129.0, 128.9, 128.3, 128.2, 126.4, 126.2, 122.8, 113.3, 112.3, 110.5, 110.2, 98.2, 59.5, 55.8, 50.7, 43.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 28 H 24 N3O5 482.1716; found: 482.1719. Characterization data of product 3s: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (minor)= 11.9 min, t R (major) = 23.2 min; 1 H NMR (400 MHz, CDCl3): δ 7.90 (d, J = 7.2 Hz, 2H), 7.54 (t, J = 6.8 Hz, 5H), 7.46-7.37 (m, 4H), 7.25 (d, J = 10.4 Hz, 1H), 6.82 (t, J = 12.4 Hz, 1H), 6.25 (s, 1H), 6.02-5.93 (m, 1H), 5.45 (d, J = 10.4 Hz, 1H), 5.36 (d, J = 16.8 Hz, 1H), 3.68-3.61 (m, 1H), 2.93 (t, J = 12.8 Hz, 1H), 2.24 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3):δ 175.1, 154.0, 142.6, 133.8, 132.9, 131.5, 130.0, 129.9, 129.4, 129.4, 129.2, 128.9, 128.8, 128.7, 128.5, 126.7, 126.6, 126.0, 125.1, 123.2, 111.2, 110.1, 98.3, 59.3, 50.4, 43.4. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 ClN3O4 486.1221; found: 486.1224. Characterization data of product 3t: The ee was determined by HPLC (Chiralpak AD-H column, i i-PrOH / hexane = 6 / 94 v / v, flow rate 0.8 mL / min, λ λ = 210 nm, 25 o °C); major diastereomer: t R (minor) = 19.5 min, t R (major) = 44.1 min; 1 ¹H NMR (400 MHz, CDCl₃): δ 7.78 (d, J J = 6.8 Hz, 2H), 7.48-7.36 (m, 5H), 7.31-7.24 (m, 3H), 7.05 (t, J J = 14.0 Hz, 1H), 6.91 (s, 1H), 6.57 (dd, J J = 8.0 Hz, 1H), 6.14 (s, 1H), 5.91-5.83 (m, 1H), 5.34 (d, J J = 10.0 Hz, 1H), 5.26 (d, J J = 16.8 Hz, 1H), 3.68-3.59 (m, 1H), 2.82 (t, J J = 12.8 Hz, 1H), 2.12 (q, J J = 12.4 Hz, 1H). 13C NMR (101MHz, CDCl3): δ 175.3, 175.1, 153.6, 145.2, 145.0, 133.4, 132.8, 132.0, 131.3,130.0, 129.0 128.9, 126.6, 126.1, 125.9, 125.8, 125.6, 123.2, 123.0, 119.0,113.5, 110.0, 98.1, 94.4, 58.9, 50.7, 43.5. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 BrN3O4 530.0715; found: 530.0713. Characterization data of product 3u: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 12 / 88 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o °C); major diastereomer: t R (minor)= 16.0 min, t R (major) = 17.3 min; 1 H NMR (400 MHz, CDCl3): δ 7.78-7.74 (m, 1H), 7.53 (q, J = 15.2 Hz, 3H), 7.44 (d, J = 6.8 Hz, 3H),7.31-7.23 (m, 3H),7.12-7.05 (m, 2H), 6.88 (d, J = 8.0 Hz, 1H), 6.20 (s, 1H),6.03-5.94 (m, 1H), 5.22 (d, J = 10.0 Hz, 1H), 5.17 (d, J = 17.2 Hz, 1H), 3.91-3.85 (m, 1H), 2.94 (t, J = 12.4 Hz, 1H), 2.30 (q, J= 13.2 Hz, 1H). 13 C NMR (101MHz, CDCl3): δ 175.8, 160.3 (d, J = 253.5 Hz, 1C), 150.0, 143.7, 134.0, 132.9 (d, J = 8.1 Hz, 1C), 132.6, 132.2, 129.8, 129.4, 128.5, 127.0, 126.7, 124.8,124.5, 123.4, 121.1, 116.8, 116.6, 114.9, 114.8, 110.8, 110.2, 97.3, 59.1,50.7, 43.4. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 FN3O4470.1516; found:470.1517. Characterization data of product 3v: The ee was determined by HPLC (Chiralpak AD-3 column, i i-PrOH / hexane = 5 / 95 v / v, flow rate 1.0 mL / min, λ = 214 nm, 25 o °C); major diastereomer: t R (major) = 16.9 min; 1 H NMR (400 MHz, CDCl3): δ 7.52 (t, J = 8.0 Hz,3H), 7.43-7.37 (m, 5H), 7.30 (t, J = 6.8 Hz, 2H), 7.22 (t, J = 7.2 Hz, 1H), 6.98(dd, J = 1.2, 1.2 Hz, 1H), 6.86 (d, J = 7.6 Hz, 1H), 5.99 (s, 1H), 5.89-5.80 (m,1H), 5.39 (d, J = 10.0 Hz, 1H), 5.29 (d,J = 17.2 Hz, 1H), 4.41-4.35 (m, 1H), 3.80 (s, 3H), 2.81 (t, J = 13.2 Hz, 1H), 2.47 (q, J = 13.6 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 174.7, 159.6, 153.8, 144.0, 134.0, 133.1, 129.8, 129.5, 128.5, 128.1, 128.1, 126.8, 123.9, 122.9, 122.7, 122.0, 117.2, 113.9, 110.9, 110.0, 99.7, 59.3, 55.6, 49.9, 43.0. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 28 H 24 N3O5 482.1716; found: 482.1719. Characterization data of product 3w: The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (minor) = 21.8 min, t R (major) = 12.0 min; 1 H NMR (400 MHz, CDCl3): δ 7.66 (t, J = 8.0 Hz, 2H), 7.55 (t, J = 8.0 Hz, 2H), 7.44 (t, J = 6.0 Hz, 4H), 7.28-7.22 (m, 2H), 7.01 (t, J = 7.6 Hz, 1H), 6.88 (d, J = 8.0 Hz, 1H), 6.84 (d, J= 7.6 Hz, 1H), 6.29 (s, 1H), 6.01-5.92 (m, 1H), 5.47 (d, J = 10.0 Hz, 1H),5.38 (d, J = 16.8 Hz, 1H), 3.77-3.70 (m, 1H), 2.95 (t, J = 12.8 Hz, 1H), 2.25 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.3, 162.9 (d, J = 247.5 Hz, 1C),152.7, 143.9 134.0, 133.0, 130.7, 130.6, 129.9, 129.5, 128.6, 126.9, 126.7,124.7, 124.7, 124.3, 123.2, 118.3, 118.1, 115.9, 115.7,110.4, 109.9, 98.5,59.2, 51.0, 43.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 FN3O4470.1516;found: 470.1518. Characterization data of product 3x: The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ = 214 nm, 25 o °C); major diastereomer: t R (minor)= 23.4 min, t R (major) = 12.2 min; 1 H NMR (400 MHz, CDCl3): δ 7.92 (s, 1H), 7.80 (d, J= 7.6 Hz, 1H), 7.57-7.50 (m, 3H), 7.44-7.41(m, 4H), 7.28-7.24 (m, 1H), 7.01 (t, J = 7.6 Hz, 1H), 6.88 (d, J = 8.0 Hz, 1H),6.83 (d, J = 7.2 Hz, 1H), 6.29 (s, 1H), 6.00-5.92 (m, 1H), 5.49 (d, J = 10.0 Hz,1H), 5.40 (d, J = 16.8 Hz, 1H), 3.77-3.70 (m, 1H), 2.95 (t, J = 12.8 Hz, 1H),2.25 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.3, 152.6, 143.9 135.1,134.0, 132.9, 131.1, 130.3, 129.9, 129.5, 128.8, 128.6, 128.3, 126.9, 126.8,126.7, 124.2, 123.3, 123.2, 110.3, 110.0, 98.5, 59.2, 51.1, 43.9.HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 ClN3O4486.1221; found: 486.1217. Characterization data of product 3y: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 5 / 95 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (minor)= 21.0 min, t R (major) = 24.9 min; 11H NMR (400 MHz, CDCl3): δ 8.07 (s, 1H), 7.85 (d, J = 7.6 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.55 (t, J = 8.0 Hz, 2H), 7.44 (t, J = 8.0 Hz, 3H), 7.37 (t, J = 8.0 Hz, 1H), 7.28-7.24 (m, 1H), 7.02 (t, J = 7.2 Hz, 1H), 6.88 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 7.6 Hz, 1H), 6.29 (s, 1H), 6.00-5.91 (m, 1H), 5.50 (d, J = 10.0 Hz, 1H), 5.41 (d, J = 16.8 Hz, 1H), 3.77-3.70 (m, 1H), 2.95 (t, J = 12.8 Hz, 1H), 2.25 (q, J = 12.8 Hz, 1H). 13 13C NMR (101 MHz, CDCl3): δ 175.3, 152.5, 143.9, 134.0, 134.0, 132.9, 131.7, 130.6, 129.8, 129.5, 128.6, 128.5, 127.3, 126.8, 126.7, 124.2, 123.4, 123.2, 123.1, 110.3, 110.0, 98.4, 59.2, 51.1, 43.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 BrN3O4 530.0715; found: 530.0714. Characterization data of product 3z: The ee was determined by HPLC (Chiralpak AD-H column, i-PrOH / hexane = 6 / 94 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o C); majordiastereomer: t R (minor)= 17.5 min, t R (major) = 34.3 min; 1 H NMR (400 MHz,CDCl3): δ 7.81 (d, J = 8.0 Hz, 2H), 7.54 (t, J = 7.6 Hz, 2H), 7.44-7.42 (m, 3H),7.33 (d, J = 8.0 Hz, 2H), 7.24 (t, J = 8.0 Hz, 1H), 7.0 (t, J = 7.6 Hz, 1H), 6.88(t, J = 8.4 Hz, 2H), 6.25 (s,1H), 6.03-5.95 (m, 1H), 5.43 (d, J = 10.0 Hz, 1H),5.35 (d, J = 16.8 Hz, 1H), 3.78-3.71 (m, 1H), 2.93 (t, J = 12.8 Hz, 1H), 2.74 (q, J = 14.8 Hz, 2H), 2.22 (q, J = 13.2 Hz, 1H), 1.30 (t, J = 7.6 Hz, 3H). 13 C NMR (101MHz, CDCl3): δ 175.6, 153.5, 147.9, 143.9,134.1, 133.3, 129.8, 129.4, 129.0,128.6, 127.1, 126.7, 124.5, 123.8, 123.1, 122.7, 110.4, 110.2, 98.2, 59.2,50.7, 43.9, 28.9, 15.3. HRMS (ESI-TOF) m / z : [M + H] + calcd forC 29H 26 N3O4480.1926; found: 480.1923. Characterization data of product 3a': The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 20 / 80 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o C); major diastereomer: t R (major)= 19.5 min; 1 H NMR (400 MHz, CDCl3): δ 7.85 (d, J = 8.8 Hz, 2H),7.55 (t, J = 8.0 Hz, 2H), 7.44 (d, J = 7.6 Hz, 3H), 7.28-7.24 (m, 1H), 7.03-7.01 (m, 3H), 6.89 (t, J = 7.2 Hz, 2H), 6.26 (s, 1H), 6.04-5.95 (m, 1H), 5.44(d, J = 10.0 Hz, 1H),5.36 (d, J = 17.2 Hz, 1H), 3.90 (s, 3H), 3.78-3.71 (m, 1H),2.94 (t, J = 12.8 Hz, 1H), 2.24 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.6, 161.9, 153.2, 143.9, 134.1, 133.3, 130.7, 129.8, 129.4, 128.6, 127.1,126.7, 124.5, 123.1, 122.7, 118.7, 114.5, 112.1, 110.5, 110.2, 98.0, 59.2,55.6, 50.7, 43.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C28 H 24 N3O5482.1719; found: 482.1716. Characterization data of product 3b': The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o °C); major diastereomer: t R (minor)= 24.3 min, t R (major) = 12.7 min; 1 H NMR (400 MHz, CDCl3): δ 7.91(q, J = 8.4 Hz, 2H), 7.55 (t, J = 7.6 Hz, 2H), 7.45 (t, J = 8.0 Hz, 3H), 7.28-7.24 (m, 1H), 7.19 (t, J = 8.8 Hz, 2H), 7.01 (t, J = 7.6 Hz, 1H), 6.87(dd, J = 13.6, 5.6 Hz, 2H), 6.28 (s, 1H), 6.01-5.92 (m, 1H), 5.44 (d, J = 10.0 Hz, 1H), 5.35 (d, J = 16.8 Hz, 1H), 3.75-3.68 (m, 1H), 2.95 (t, J = 12.8 Hz, 1H), 2.24 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.4, 164.4 (d, J= 254.5 Hz, 1C), 152.6, 144.0, 134.0, 133.1, 131.2, 131.1, 129.8, 129.5, 128.6, 126.9, 126.7, 124.3, 123.1, 123.0, 122.7, 116.4, 116.2, 110.3, 110.2, 98.3, 59.2, 50.9, 43.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 FN3O4 470.1516; found: 470.1517. Characterization data of product 3c': The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 6 / 94 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o °C); major diastereomer: t R (minor) = 29.4 min, t R (major) = 15.1 min; 1 ¹H NMR (400 MHz, CDCl₃): δ 7.75 (d, J = 8.4 Hz, 2H), 7.43 (q, J = 15.2 Hz, 2H), 7.38-7.32 (m, 5H), 7.15 (t, J = 7.6 Hz, 1H), 6.90 (t, J = 6.8 Hz, 1H), 6.78 (d, J = 8.0 Hz, 1H), 6.73 (d, J = 7.2 Hz, 1H), 6.18 (s, 1H), 5.90-5.81 (m, 1H), 5.35 (d, J = 10.0 Hz, 1H), 5.26 (d, J = 16.8 Hz, 1H), 3.64-3.57 (m, 1H), 2.84 (t, J= 13.2 Hz, 1H), 2.14 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl₃): δ 175.3, 152.7, 143.9, 137.4, 134.0,133.0, 130.1, 129.8, 129.6, 129.5, 129.3, 128.6, 126.8, 126.7, 126.7, 125.0,124.2, 123.1, 110.3, 110.0, 98.4, 59.1, 50.9, 43.8. HRMS (ESI-TOF) m / z : [M +H] + calcd for C 27 H 21 ClN₃O₄ 486.1221; found: 486.1223. Characterization data of product 3d': The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 6 / 94 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o °C); major diastereomer: t R (minor)= 18.7 min, t R (major) = 40.0 min; 1 H NMR (400 MHz, CDCl₃): δ 7.78 (d, J = 8.8 Hz, 2H), 7.64 (d, J = 8.8 Hz, 2H), 7.55 (t, J = 8.0 Hz, 2H), 7.45 (t, J = 7.6 Hz, 3H), 7.28-7.24 (m, 1H), 7.00 (t, J = 7.2 Hz, 1H), 6.89 (d, J = 8.0 Hz, 1H), 6.82 (d, J= 7.6 Hz, 1H), 6.29 (s, 1H), 6.00-5.91 (m, 1H),5.45 (d, J = 10.0 Hz, 1H), 5.36 (d, J = 16.8 Hz, 1H), 3.74-3.67 (m, 1H), 2.94 (t, J = 12.8 Hz, 1H), 2.24 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.3,152.8, 143.9, 134.0, 133.0, 132.3, 130.3, 129.8, 129.5, 128.6, 126.9, 126.7,125.8, 125.5, 124.2, 123.1, 123.1, 110.3, 110.0, 98.4, 59.2, 51.0, 43.9. HRMS(ESI-TOF) m / z : [M + H] + calcd for C 27 H 21 BrN3O4530.0715; found: 530.0718. Characterization data of product 3e': The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 3 / 97 v / v, flow rate 1.0 mL / min, λ = 254 nm, 25 o C); major diastereomer: t R (major) = 38.5 min; 1 H NMR (400 MHz, CDCl3): δ 8.06 (d, J = 8.0 Hz,2H), 7.78 (d, J = 8.0 Hz, 2H), 7.56 (t, J = 8.0 Hz, 2H), 7.46 (t, J = 6.4 Hz, 3H),7.29 (d, J= 7.6 Hz, 1H), 7.02 (t, J = 7.6 Hz, 1H), 6.90 (d, J = 8.0 Hz, 1H), 6.81(d, J = 7.6 Hz, 1H), 6.34 (s, 1H), 6.03-5.94 (m, 1H), 5.50 (d, J = 10.0 Hz, 1H),5.40 (d, J = 16.8 Hz, 1H), 3.77-3.70 (m, 1H), 2.98 (t, J = 12.8 Hz, 1H), 2.27 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.3, 152.6, 144.0, 134.0, 133.0,133.0, 132.6, 132.3, 130.2, 129.9, 129.6, 129.2, 128.7, 126.8, 126.7, 126.0(q, J = 4.0 Hz, 2C), 123.7 (q, J = 273.7 Hz, 1C), 123.2, 110.5, 110.0, 98.7,59.2, 51.2, 43.9. HRMS (ESI-TOF) m / z : [M + H] + calcd for C 28 H 21 F3N3O4520.1484;found: 520.1486. Characterization data of product 3f': The ee was determined by HPLC (Chiralpak IH column, i -PrOH / hexane = 20 / 80 v / v, flow rate 0.8 mL / min, λ = 254 nm, 25 o °C); major diastereomer: t R (major)= 31.8 min; 1 H NMR (400 MHz, CDCl3): δ8.16 (d, J = 8.4 Hz, 2H), 7.98 (d, J = 8.4 Hz, 2H), 7.54 (q, J = 15.6 Hz, 2H), 7.46-7.43 (m, 3H),7.28-7.24 (m, 1H), 7.0 (t, J = 7.2 Hz, 1H), 6.89 (d, J = 7.6 Hz, 1H), 6.82 (d, J =7.6 Hz, 1H), 6.31 (s, 1H), 6.02-5.93 (m, 1H), 5.47 (d, J = 10.4 Hz, 1H), 5.37(d, J = 17.2 Hz, 1H), 3.97 (s, 3H), 3.77-3.70 (m, 1H), 2.96 (t, J = 12.8 Hz, 1H),2.25 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.3, 166.3, 152.9, 143.9,134.0, 133.0, 132.3, 130.7, 130.1, 130.1, 129.8, 129.5, 128.8, 128.6, 126.8,126.7, 124.2, 123.2, 123.1, 110.3, 110.0, 98.6, 59.2, 52.5, 51.0, 43.9. HRMS(ESI-TOF) m / z : [M + H] + calcd for C 29 H 24 N3O6510.1667; found: 510.1665. Characterization data of product 3g': The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 6 / 94 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 oC); majordiastereomer: t R (major) = 25.3 min; 1 H NMR (400 MHz, CDCl3): δ 7.97 (d, J = 8.0 Hz, 2H), 7.73 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 7.6 Hz, 2H), 7.57-7.49 (m, 3H), 7.47-7.39 (m,5H), 7.28-7.24 (m, 1H), 7.02 (t, J = 7.6 Hz, 1H), 6.91 (d, J = 8.0 Hz, 1H), 6.88 (d, J =8.0 Hz, 1H), 6.30 (s, 1H), 6.06-5.98 (m, 1H), 5.46 (d, J = 10.0Hz, 1H), 5.40 (d, J = 16.8 Hz, 1H), 3.82-3.75 (m, 1H), 2.96 (t, J = 12.8 Hz, 1H), 2.25 (q, J = 12.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.5, 153.4, 144.0, 143.9,140.0, 140.0, 139.9, 134.1, 133.3, 129.9, 129.4, 129.1, 128.6, 128.3, 127.7,127.3, 127.1, 126.8, 125.3, 124.5, 123.2, 122.9, 110.3, 98.3, 59.3, 50.9,44.0. HRMS (ESI-TOF) m / z [M + H] + calcd for C 33 H 26 N3O4528.1926; found: 528.1923. Product characterization data at 3h: The eewas determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 20 / 80 v / v, flow rate 0.4 mL / min, λ = 254 nm, 25 o C); majordiastereomer: t R (minor)= 24.8 min, t R (major) = 33.5 min; 1 H NMR (400 MHz,CDCl3): δ 7.97 (d, J = 2.0 Hz, 1H), 7.79 (dd, J = 8.4, 2.0 Hz, 1H), 7.54 (q, J = 15.6Hz, 2H), 7.45 (t, J = 4.4 Hz, 3H), 7.28-7.24 (m, 1H), 7.03 (dd, J = 14.4, 8.4 Hz,2H), 6.87 (dd, J = 13.2, 8.0 Hz, 2H), 6.27 (s, 1H), 6.00-5.91 (m, 1H), 5.49 (d, J = 10.0 Hz, 1H), 5.41 (d, J = 16.8 Hz, 1H), 3.99 (s, 3H), 3.77-3.70 (m, 1H),2.93 (t, J = 13.2 Hz, 1H), 2.24 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.4, 157.2, 152.3, 143.9, 134.0, 133.0, 130.6, 129.8, 129.4, 128.8, 128.6,126.9, 126.7, 124.3, 123.3, 123.2, 123.1, 119.6, 112.1, 110.3, 110.2, 98.1,59.2, 56.4, 50.9, 43.9. HRMS (ESI-TOF) m / z[M + H] + calcd forC 28 H 23 ClN3O5516.1326; found: 516.1331. Example 54: ; In a dry 10 mL reaction tube, first add 0.10 mmol of 3-phenyl-4-nitroisoxazole 1a and 0.001 mmol of Rh2(OAc)4, then add 1.0 mL of dichloromethane. The mixture is then incubated at 25 °C. o Activation was performed for 1 h under C and argon protection with stirring. Then, 0.12 mmol of spironindione-vinylcyclopropane 4, 0.005 mmol of Pd2(dba)3, and 0.01 mmol of chiral bisphosphine ligand L6 were added sequentially to the above reaction solution, followed by 2.0 mL of dichloromethane to completely dissolve the ligands. The mixture was then placed at -10°C. o The mixture was stirred for 48 h under the protection of C and argon. The reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was separated and purified by column chromatography to obtain chiral compound 5 (80% yield, 64% ee, >20:1 dr).

[0023] Product 5 characterization data: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ = 254 nm, 25 o C); majordiastereomer: t R (major) = 20.1 min; 1 H NMR (400 MHz, CDCl3): δ 7.93 (d, J = 23.6Hz, 2H), 7.82 (s, 2H), 7.76 (d, J = 6.8 Hz, 2H), 7.37 (q, J = 7.2 Hz, 3H), 5.92(s, 1H), 5.78-5.69 (m, 1H), 5.28 (d, J = 10.0 Hz, 1H), 5.15 (d, J= 16.8 Hz, 1H),4.00-3.94 (m, 1H), 2.64 (t, J = 12.8 Hz, 1H), 2.07 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 198.3, 198.0, 154.0, 141.9, 141.8, 136.7, 136.5, 132.8,130.9, 129.3, 128.7, 126.5, 124.0, 123.9, 123.0, 110.6, 96.5, 63.5, 50.3,39.8, 29.8. HRMS (ESI-TOF) m / z [M + H] + calcd for C 22 H 17 N2O5 389.1137; found: 389.1127. Example 55:

[0024] In a dry 10 mL reaction tube, first add 0.10 mmol of 3-phenyl-4-nitroisoxazole 1a and 0.001 mmol of Rh2(OAc)4, then add 1.0 mL of dichloromethane. The mixture is then incubated at 25 °C. o Activation was performed for 1 h under C and argon protection with stirring. Then, 0.12 mmol of spirobenzofuran-3-one-vinylcyclopropane 6, 0.005 mmol of Pd2(dba)3, and 0.01 mmol of chiral bisphosphine ligand L6 were added sequentially to the above reaction solution, followed by 2.0 mL of dichloromethane to completely dissolve the ligands. The mixture was then placed at -10°C. o The mixture was stirred for 48 h under C and argon protection. The reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain chiral compound 7 (30% yield, 99% ee, 5:1 dr).

[0025] Characterization data of product 7: The ee was determined by HPLC (Chiralpak AD-H column, i -PrOH / hexane = 5 / 95 v / v, flow rate 1.0 mL / min, λ = 210 nm, 25 oC); majordiastereomer: t R (major) = 21.2 min; 1 H NMR (400 MHz, CDCl3): δ 7.80 (d, J = 7.2 Hz, 2H), 7.61 (t, J = 6.8 Hz, 2H), 7.43-7.37 (m, 2H), 7.20 (s, 1H), 7.09 (dd, J =14.0, 8.4 Hz, 2H), 5.98 (s, 1H), 5.83-5.74 (m, 1H), 5.31 (d, J = 10.0 Hz, 1H), 5.19 (d, J = 16.8 Hz, 1H), 3.92 (q, J = 19.2 Hz, 1H), 2.69 (t, J = 12.4 Hz, 1H), 2.20 (q, J = 13.2 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 188.6, 178.2, 153.9, 138.8,132.7, 131.0, 129.4, 128.8, 125.1, 123.0, 122.9, 120.0, 116.6, 113.5, 108.8,96.0, 94.8, 47.3, 39.0. HRMS (ESI-TOF) m / z [M + H] + calcd forC 21 H 17 N2O5377.1137; found: 377.1140. Example 56:

[0026] In a dry 10 mL reaction tube, first add 0.10 mmol of 3-phenyl-4-nitroisoxazole 1a and 0.001 mmol of Rh2(OAc)4, then add 1.0 mL of dichloromethane. The mixture is then incubated at 25 °C. oActivation was performed for 1 h under C and argon protection with stirring. Then, 0.12 mmol of spiroisoquinoline-1,3-diketone-vinylcyclopropane 8, 0.005 mmol of Pd2(dba)3, and 0.01 mmol of chiral bisphosphine ligand L4 were added sequentially to the above reaction solution, followed by 2.0 mL of dichloromethane to completely dissolve the ligands. The mixture was then placed at -10°C. o The mixture was stirred for 48 h under the protection of C and argon. The reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain chiral compound 9 (65% yield, 60% ee, >20:1 dr).

[0027] Characterization data of product 9: The ee was determined by HPLC (Chiralpak OD-H column, i -PrOH / hexane = 10 / 90 v / v, flow rate 0.8 mL / min, λ = 210 nm, 25 o C); majordiastereomer: t R (minor) = 19.6 min, t R (major) = 12.9 min; 1 H NMR (400 MHz, CDCl3): δ 8.29 (d, J = 8.0 Hz, 1H), 7.82 (t, J = 7.2 Hz, 4H), 7.57-7.54 (m, 1H), 7.48-7.41 (m, 3H), 5.92 (s, 1H), 5.87-5.79 (m, 1H), 5.40 (d, J = 10.0 Hz, 1H), 5.29 (d, J = 16.8 Hz, 1H), 4.47 (q, J = 19.2 Hz, 1H), 3.37 (s, 3H), 2.90 (t, J =13.6 Hz, 1H), 2.71 (q, J = 13.6 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ171.8, 163.6,154.4, 139.1, 135.1, 133.4, 130.8, 129.4, 129.2, 128.9,128.6, 126.6, 125.5,125.3, 123.2, 112.5, 104.3, 59.0, 52.1, 48.5, 27.5. HRMS (ESI-TOF) m / z [M +H] + calcd for C 23 H 20 N3O5418.1403; found: 418.1407. Antibacterial activity verification: The chiral spirocyclopentane-isoxazoline compounds prepared in the examples were selected, and their activity in inhibiting lipopolysaccharide-induced NO release from RAW264.7 macrophages was detected using the Griess method to investigate their anti-inflammatory activity and applications.

[0028] The following description uses compound 3a from Example 20 as an example. The specific experimental procedures are as follows: Experimental methods: 1. Culture of macrophage line RAW264.7 Prepare a solution containing 10% fetal bovine serum and 100 U / mL of DMEM medium as the base. -1 Penicillin and 100 U·mL -1 Cell culture medium containing streptomycin and 50 μM 2-mercaptoethanol. Macrophages were cultured at approximately 4 × 10⁻⁶ cells / mL. 5 cells·mL -1 The cells were passaged in 5% CO2 culture flasks at 37 °C. By day 3, adherent cells covered approximately 50%–60% of the bottom area of ​​the culture flask. The adherent cells were then digested with trypsin and passaged to another culture flask. RAW cells that had been cryopreserved and thawed at -80 °C were used as the first generation, and RAW264.7 cells from the 3rd to 8th generations were selected for experiments.

[0029] 2. Griess method for detecting the inhibitory effect of compounds on LPS-activated macrophages RAW264.7 macrophages in logarithmic growth phase were harvested and their density adjusted to 1 × 10⁵ cells / mL using fresh DMEM medium containing 10% fetal bovine serum. -1Cells were seeded in 96-well plates at 100 μL / well and cultured at 37 ℃ in a 5% CO2 incubator. After 24 h of adherent culture, the medium was replaced with serum-free fresh medium, and the cells were simultaneously treated with the drug. Sample 3a was treated with lipopolysaccharide at doses of 0.1, 0.5, 2.5, 12.5, and 62.5 μM. A blank control group was also included. The final concentration of LPS in each treatment group and the positive control group was 10 ng / mL. -1 After cell culture for 24 hours following drug administration, the supernatant was collected, and the NO content in the supernatant was detected using the Griess colorimetric method. 2- content.

[0030] 3. Statistical Methods All data were analyzed using SPSS (13.0) statistical software. Results are expressed as mean ± standard error. To assess overall differences, one-way ANOVA was used to analyze homogeneity of variance between groups, combined with Dunnett's test for inter-group comparisons. The Levene test was used to test homogeneity of variance in multiple samples. When p > 0.05, the variances were homogeneous, and Dunnett's two-tailed t-test was used to assess the difference in means among multiple groups. When p < 0.05, the variances were unequal, and Dunnett's T3 test was used to assess the difference in means among multiple groups. See Table 3 for details.

[0031]

[0032] The results showed that the chiral cyclopentane-isoxazoline compounds prepared in this invention exhibited varying degrees of inhibition of lipopolysaccharide-induced NO release from RAW264.7 macrophages. Among them, compounds 3d (IC50: 27.54 ± 0.89), 3g (IC50: 18.59 ± 0.62), 3i (IC50: 3.87 ± 0.13), 3w (IC50: 6.32 ± 0.12), 3x (IC50: 11.56 ± 0.16), 5 (IC50: 12.65 ± 1.08), and 7 (IC50: 4.28 ± 0.17) showed particularly superior inhibitory activity, especially compounds 3i, 3w, and 7. The results of the above study indicate that the chiral products 3i, 3w, and 7 are highly active potential anti-inflammatory compounds obtained in this screening, providing experimental evidence for the development of novel anti-inflammatory drugs. Further structural optimization can enhance their anti-inflammatory application potential.

[0033] Compounds 3a, 3c, and 9 were extracted and analyzed by nuclear magnetic resonance spectroscopy and chiral high-performance liquid chromatography. Compounds 3j and 5 were extracted and analyzed for single-crystal structure.Figure 1 to Figure 14 As shown.

[0034] The above descriptions are merely embodiments of the present invention, and common knowledge such as specific technical solutions and / or characteristics are not described in detail here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A chiral spirocyclopentane-isoxazoline compound, characterized in that, Its general structural formula is as follows: ， The spirocyclic skeleton in the formula can be independently represented as a benzo5-membered nitrogen heterocycle, a benzo5-membered carbon ring, a benzo5-membered oxygen heterocycle, or a benzo6-membered nitrogen heterocycle.

2. The chiral spirocyclopentane-isoxazoline compound according to claim 1, characterized in that: (1) When the spirocyclic skeleton is a benzo5-membered nitrogen heterocycle, the general formula of the compound is as follows: ， Among them, Ar 1 It is independently selected from phenyl, 2-fluorophenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-ethylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-trifluoromethylphenyl, 4-methoxycarbonylphenyl, 4-phenylphenyl or 3-chloro-4-methoxyphenyl; Accordingly, Ar 2 It is independently selected from phenyl, 4-chlorophenyl, 4-bromophenyl, 5-methylphenyl, 5-methoxyphenyl, 5-chlorophenyl, 5-bromophenyl, 6-chlorophenyl, 6-bromophenyl, 7-methylphenyl, 7-chlorophenyl, 7-bromophenyl, 4-bromo-5-methylphenyl, 5,7-dimethylphenyl or 5-bromo-7-methylphenyl; Accordingly, R is independently selected from methyl, benzyl, or phenyl; (2) When the spirocyclic skeleton is a benzo5-membered carbon ring, the general formula of the compound is as follows: ； (3) When the spirocyclic skeleton is a benzo5-membered oxygen heterocycle, the general formula of the compound is as follows: ； (4) When the spirocyclic skeleton is a benzo-6-membered nitrogen heterocycle, the general formula of the compound is as follows: 。 3. The chiral spirocyclopentane-isooxazoline compound according to claim 2, characterized in that: Its structural formula is as follows: ， 。 4. The use of the chiral spirocyclopentane-isoxazolinoid compound according to any one of claims 1 to 3 in the preparation of anti-inflammatory drugs, characterized in that: The anti-inflammatory effect is achieved by inhibiting the release of the inflammatory factor NO from RAW264.7 macrophages induced by lipopolysaccharide.

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