A chiral spiro [oxindole-pyrrolidine amino acid ester] compound containing trifluoromethyl and a preparation method and application thereof

Abstract

This invention discloses a chiral spiro[oxoindole-pyrrolidine amino acid ester] compound containing trifluoromethyl groups in the field of organic synthesis. Using indigo-derived trifluoromethylimine ylide and 2-amidoacrylate substrates with good activity, a highly stereoselective one-step asymmetric [3+2] cyclization reaction catalyzed by inexpensive metal / chiral ligand complexes and a base is employed to obtain the chiral spiro[oxoindole-pyrrolidine amino acid ester] compound containing trifluoromethyl groups. This invention effectively enriches the asymmetric synthesis technology and molecular structural diversity of chiral spirocyclic pyrrolidine amino acid derivatives, and is of great significance for in-depth drug activity research based on new drug development.

Description

Technical Field

[0001] This invention belongs to the field of organic synthesis technology, and specifically relates to a chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl groups and its preparation method. Background Technology

[0002] Spirotry[indole-pyrrolidine] and pyrrolidine amino acid derivatives are two very important classes of heterocyclic building blocks, widely found in pharmaceutical active molecules and natural products, such as the natural products Spirotryprostatin A and (S)-(-)-Cucurbitine. These molecular structures typically possess superior antiviral, antibacterial, antiparasitic, and anti-neurological drug activities, making them important lead compounds for new drug development. Therefore, the synthesis of these compounds has attracted widespread attention from synthetic scientists.

[0003] Current technical approaches mainly focus on the asymmetric synthesis of spiro[indole-pyrrolidine] skeletons. For chiral skeletons of pyrrolidine amino acid derivatives, the main approach is to first obtain the racemic mixture through natural extraction or total synthesis, and then resolve it. However, the asymmetric synthesis of molecules with both spiro[indole-pyrrolidine] and pyrrolidine amino acid dual pharmacophore structures remains a gap and a challenging task. Summary of the Invention

[0004] This invention addresses the shortcomings of existing technologies by providing a chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl groups and its preparation method. The main purpose is to enrich the asymmetric synthesis technology and molecular structural diversity of chiral spirocyclic pyrrolidine amino acid derivative compounds, laying a material foundation for in-depth drug activity research based on new drug development.

[0005] The general structural formula of a chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl in this scheme is as follows:

[0006]

[0007] Ar 1 It is selected from any one of phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 5-methylphenyl, 5-methoxyphenyl, 5-fluorophenyl, 5-chlorophenyl, 5-bromophenyl, 5-nitrophenyl, 6-methoxyphenyl, 6-fluorophenyl, 6-chlorophenyl, 6-bromophenyl, 7-methylphenyl, 7-fluorophenyl, 7-chlorophenyl, 7-bromophenyl, 7-trifluoromethylphenyl, 5,7-dimethylphenyl, 4-pyridyl, and 7-pyridyl.

[0008] R 1It is selected from any one of hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, allyl, benzyl, and phenyl;

[0009] R 2 It is selected from any one of hydrogen, formyl, acetyl, benzoyl, tert-butoxycarbonyl, p-valeryl, benzyloxycarbonyl, methanesulfonyl, and p-toluenesulfonyl.

[0010] R 3 It is selected from any one of hydrogen, formyl, acetyl, benzoyl, tert-butoxycarbonyl, p-valeryl, benzyloxycarbonyl, methanesulfonyl, and p-toluenesulfonyl.

[0011] Or, R 2 and R 3 All are phthaloyl groups;

[0012] R is selected from any one of methyl, ethyl, tert-butyl, benzyl, and phenyl.

[0013] The present invention also provides a method for preparing the chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl, comprising the following steps:

[0014] Step 1: Add the metal catalyst, chiral ligand, base, indigo-derived trifluoromethylimine ylide and 2-amidoacrylate to the reaction tube in sequence, then add the organic solvent, and stir the mixture at a temperature of 0-50℃.

[0015] Step 2: After the reaction is complete, the chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl is obtained by post-processing and separation.

[0016] The synthesis route is shown below:

[0017]

[0018] The principle of this method is as follows: a metal / chiral ligand and a base catalyze the asymmetric [3+2] cyclization reaction of indigo-derived trifluoromethyleneimine ylide and 2-amidoacrylate substrates via the following mechanism (using cuprous chloride / L3 and cesium carbonate catalysis as an example). First, the indigo-derived trifluoromethyleneimine ylide is deprotonated by cesium carbonate to generate a trifluoromethyleneimine ylide anion. Next, this anion further coordinates with the in-situ generated chiral metal complex. Under this chiral induction, the Si facet of the coordinated trifluoromethyleneimine ylide anion attacks the Si facet of the 2-amidoacrylate, completing a stereoselective [3+2] cyclization tandem reaction to generate a chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing a trifluoromethyl group. The principle of the reaction is shown in the following equation:

[0019]

[0020] The beneficial technical effects of this method are as follows: This invention uses indigo-derived trifluoromethylimine ylide and 2-amidoacrylate substrates with good activity, and obtains chiral spiro[oxyindole-pyrrolidine amino acid ester] compounds containing trifluoromethyl in one step with high stereoselectivity through a low-cost metal / chiral ligand complex and base-catalyzed asymmetric [3+2] cyclization reaction. This effectively enriches the asymmetric synthesis technology and molecular structure diversity of chiral spirocyclic pyrrolidine amino acid derivatives, and is of great significance for in-depth drug activity research based on new drug development.

[0021] In the method of the present invention, in order to prevent the evaporation of organic solvents and the influence of moisture, step one is carried out in a sealed container.

[0022] Furthermore, the metal catalyst is a copper salt or a silver salt.

[0023] Further, the copper salt is any one of copper sulfate, basic copper carbonate, copper acetate, copper trifluoroacetate, cuprous chloride, cuprous bromide, copper chloride, copper bromide, copper cyanide, copper trifluoromethanesulfonate, cuprous oxide, and copper oxide; the silver salt is any one of silver acetate, silver nitrate, silver trifluoroacetate, silver trifluoromethanesulfonate, silver hexafluoroantimonate, and silver benzoate. Preferably, it is a metallic copper salt, more preferably cuprous chloride, as the yield and stereoselectivity of the product obtained using these salts are relatively high.

[0024] Furthermore, the chiral ligand can be any one of the following structures, preferably L3, as the product obtained using it has relatively high stereoselectivity.

[0025]

[0026] Furthermore, the organic solvent is a hydrocarbon, ether, ester, amide, nitrile, sulfone, or sulfoxide. A hydrocarbon is preferred, with toluene being particularly desirable, as the yield and stereoselectivity of the product obtained using this solvent are relatively high.

[0027] Furthermore, the base is any one of triethylamine, diisopropylethylamine, 1,8-diazabicycloundec-7-ene, sodium acetate, potassium acetate, sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, potassium phosphate, cesium tert-valerate, sodium methoxide, sodium tert-butoxide, and potassium tert-butoxide. Cesium carbonate is preferred, as the yield and stereoselectivity of the product obtained using it are relatively high.

[0028] Furthermore, the temperature described in step one is 10°C. At this temperature, the yield and stereoselectivity of the product are relatively high.

[0029] In step two of this invention, the product is separated using simple column chromatography or recrystallization. Combined with the aforementioned preferred conditions, the resulting trifluoromethyl-containing chiral spiro[oxyindole-pyrrolidine amino acid ester] compound exhibits an optical purity ≥94% (high dr and ee). This trifluoromethyl-containing chiral spiro[oxyindole-pyrrolidine amino acid ester] compound can be used in the preparation of antitumor drugs, showing significant inhibitory effects on the proliferation of human lung cancer cells A549, liver cancer cells HepG2, and cervical cancer cells HeLa. Attached Figure Description

[0030] Figure 1 Compound 3a of the present invention 1 HNMR spectrum;

[0031] Figure 2 Compound 3a of the present invention 13 C NMR spectrum;

[0032] Figure 3 Compound 3b of the present invention 1 H NMR spectrum;

[0033] Figure 4 Compound 3b of the present invention 13 C NMR spectrum;

[0034] Figure 5 Compound 3c of the present invention 1 H NMR spectrum;

[0035] Figure 6 Compound 3c of the present invention 13 C NMR spectrum;

[0036] Figure 7 The attached diagram shows a single crystal of 3g of the compound of this invention. Detailed Implementation

[0037] The present invention will be further described in detail below through specific embodiments:

[0038] This invention relates to a chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl groups and its preparation method, comprising:

[0039] Step 1: Add the metal catalyst, chiral ligand, base, indigo-derived trifluoromethylimine ylide and 2-amidoacrylate to the reaction tube in sequence, then add the organic solvent, and stir the mixture at a reaction temperature of 0℃~50℃.

[0040] Step 2: After the reaction is complete, the chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl is obtained by post-processing and separation.

[0041] For example, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs2CO3, 0.1 mmol indigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene and stirred at 10 °C for the appropriate time. Then, the reaction solution was directly separated by column chromatography to obtain the corresponding trifluoromethyl-containing chiral spiro[oxyindole-pyrrolidine amino acid ester] compound 3.

[0042] The compounds and their preparation methods of this invention utilize trifluoromethylimine ylides with different structures and 2-amidoacrylates to synthesize different compounds 3a to 3k, with the specific structural formulas as follows:

[0043]

[0044] In this invention, the diastereoselectivity, enantioselectivity, and absolute configuration of the product were measured by nuclear magnetic resonance, chiral high-performance liquid chromatography, and X-ray single-crystal diffraction, respectively.

[0045] Example 1:

[0046]

[0047] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol indigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3a (yield 92%, ee 99%, dr value > 20:1).

[0048] Characterization data of product 3a: Light yellow oil; 92% yield; >20:1 dr, 99% ee; [α] D 25 =124.8(c 0.50,CH2Cl2).The ee was determined by HPLC(Chiralpak OX-3, i-PrOH / hexane=20 / 80, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =12.0min,t major =13.1min). 1H NMR (400MHz, CDCl3): δ7.41-7.37(m,1H),7.34-7.31(m,3H),7.29-7.26(m,3H),7.06(t,J=7.6Hz,1H),6.84(d,J=8.0Hz,1H),5.44(s,1H),5.01(d d,J=15.2,12.0Hz,2H),4.30-4.22(m,1H),3.57(s,3H),3.48(dd,J=13.6 ,10.4Hz,1H),3.12(s,3H),2.92(q,J=7.2Hz,1H),2.53(d,J=8.4Hz,1H); 13 C NMR (101MHz, CDCl3): δ175.4,169.4,155.4,144.9,135.7,131.1,128.7,128.6,128.5,125.3,124.0 ,123.0,109.2,72.6,70.2,67.5,58.0(q,J=32.3Hz,1C),52.9,33.6,26.4.HRMS(ESI-TOF)calcd.for C 23 H 23 F3N3O5[M+H] + 478.1584; found: 478.1590.

[0049] Example 2:

[0050]

[0051] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 4-bromoindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3b (yield 70%, ee 98%, dr value > 20:1).

[0052] Product 3b characterization data: Light yellow oil; 70% yield; >20:1 dr, 98% ee; [α] D 25=106.9(c 0.50,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3, i-PrOH / hexane=30 / 70, flow rate 0.9mL / min, λ=214nm, major diastereomer:t minor =13.8min,t major =9.1min). 1 H NMR (400MHz, CDCl3): δ7.37-7.28(m,5H),7.27-7.20(m,2H),6.82-6.77(m,1H),5.95(s,1H),5.04(dd,J=20.0,12. 4Hz,2H),4.38-4.23(m,1H),3.58(s,3H),3.40-3.34(m,1H),3.08(s,3H),2.97(d,J=7.2Hz,1H),2.90-2.84(m,1H); 13 C NMR (101MHz, CDCl3): δ175.4,168.9,154.8,147.6,135.9,132.6,128.6,128.4,128.3,127.9,122.6 ,118.9,108.4,74.8,72.9,67.3,58.3(q,J=32.3Hz,1C),53.0,32.6,26.6.HRMS(ESI-TOF)calcd.for C 23 H 22 BrF3N3O5[M+H] + 556.0689; found: 556.0698.

[0053] Example 3:

[0054]

[0055] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 5-methoxyindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3c (yield 92%, ee 98%, dr value > 20:1).

[0056] Product 3c characterization data: Light yellow oil; 92% yield; >20:1 dr, 98% ee; [α] D 25 = 57.8 (c 0.50, CH2Cl2). The ee was determined by HPLC (Chiralpak OZ-3, i-PrOH / hexane = 20 / 80, flow rate 1.0 mL / min, λ = 254 nm, major diastereomer: t minor = 12.3 min, t major = 9.2 min). 1 1H NMR (400 MHz, CDCl3): δ 7.34 - 7.24 (m, 5H), 6.95 (d, J = 2.8 Hz, 1H), 6.89 - 6.86 (m, 1H), 6.74 (d, J = 8.4 Hz, 1H), 5.66 (s, 1H), 5.02 - 4.95 (m, 2H), 4.33 - 4.25 (m, 1H), 3.74 (s, 3H), 3.55 - 3.45 (m, 4H), 3.10 (s, 3H), 2.90 (q, J = 7.6 Hz, 1H), 2.57 (d, J = 8.4 Hz, 1H); 13 13C NMR (1z01 MHz, CDCl3): δ 175.1, 169.3, 156.1, 155.5, 138.3, 135.6, 128.7, 128.5, 128.5, 126.6, 114.4, 112.2, 109.5, 73.0, 70.4, 67.6, 58.0 (q, J = 32.3 Hz, 1C), 55.9, 52.9, 31.7, 26.5. HRMS (ESI-TOF) calcd. for C 24 H 25 F3N3O6 [M + H] + 508.1690; found: 508.1695.

[0057] Example 4:

[0058]

[0059] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 5-fluoroindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 11 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3d (yield 99%, ee 99%, dr value > 20:1).

[0060] Product 3D characterization data: Light yellow oil; 99% yield; >20:1 dr, 99% ee; [α] D 25 =46.9(c 0.50,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3, i-PrOH / hexane=20 / 80, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =6.7min,t major =7.1min). 1 H NMR (400MHz, CDCl3): δ7.34-7.25(m,5H),7.11-7.08(m,2H),6.76(q,J=4.4Hz,1H),5.53(s,1H),5.00(dd,J=18.8,12.0 Hz,2H),4.31-4.23(m,1H),3.57(s,3H),3.50-3.44(m,1H),3.12(s,3H),2.81(q,J=6.8Hz,1H),2.55(d,J=10.0Hz,1H); 13 C NMR (101MHz, CDCl3): δ175.4,169.1,159.0(d,J=243.4Hz,1C),155.3,140.9(d, J=2.0Hz,1C),135.5,128.7,128.6,128.5,126.8(d,J=7.1Hz,1C),117.1(d,J=24 .2Hz,1C),112.6(d,J=25.3Hz,1C),109.6(d,J=8.1Hz,1C),72.6(d,J=1.0Hz,1C) ,70.8,67.7,57.9(q,J=32.3Hz,1C),53.0,34.1,26.6.HRMS(ESI-TOF)calcd.for C 23 H22 F4N3O5[M+H] + 496.1490; found: 496.1495.

[0061] Example 5:

[0062]

[0063] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 5-bromoindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 11 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3e (yield 99%, ee 97%, dr value > 20:1).

[0064] Product 3e characterization data: Light yellow oil; 99% yield; >20:1 dr, 97% ee; [α] D 25 =0.4(c0.50,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3, i-PrOH / hexane=12 / 88, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =10.7min,t major =9.1min). 1 H NMR (400MHz, CDCl3): δ7.52-7.46(m,2H),7.35-7.27(m,5H),6.70(d,J=8.4Hz,1H),5.67(s,1H),5.02-4.95(m,2 H),4.36-4.25(m,1H),3.57(s,3H),3.47-3.40(m,1H),3.13(s,3H),2.76(q,J=6.8Hz,1H),2.53(d,J=8.0Hz,1H); 13C NMR (101MHz, CDCl3): δ175.3,168.9,155.2,144.1,135.4,133.6,128.7,128.6,128.6,127.6,127.4 ,115.1,110.4,72.4,71.0,67.9,57.9(q,J=32.3Hz,1C),53.0,34.2,26.6.HRMS(ESI-TOF)calcd.for C 23 H 22 BrF3N3O5[M+H] + 556.0689; found: 556.0692.

[0065] Example 6:

[0066]

[0067] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 6-chloroindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3f (yield 56%, ee 98%, dr value >20:1).

[0068] Product 3F characterization data: Light yellow oil; 56% yield; >20:1 dr, 98% ee; [α] D 25 =-41.8(c 0.27,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3, i-PrOH / hexane=20 / 80, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =7.5min,t major =6.7min). 1H NMR (400MHz, CDCl3): δ7.36-7.33(m,3H),7.27-7.25(m,2H),7.16(d,J=8.0Hz,1H),6.92-6.89(m,1H),6.81(d,J=1.6Hz,1H),5.49(s,1H),4.99(d d,J=26.8,12.0Hz,2H),4.30-4.22(m,1H),3.60(s,3H),3.44(dd,J=13.6 ,10.4Hz,1H),3.10(s,3H),2.82(q,J=7.2Hz,1H),2.51(d,J=8.0Hz,1H); 13 C NMR (101MHz, CDCl3): δ175.6,169.1,155.2,146.1,136.9,135.6,128.7,128.7,125.0,123.7,12 2.7,109.8,72.2,70.6,67.7,57.8(q,J=32.3Hz,1C),53.0,33.9,26.6.HRMS(ESI-TOF)calcd.for C 23 H 22 ClF3N3O5[M+H] + 512.1195; found: 512.1203.

[0069] Example 7:

[0070]

[0071] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 7-methylindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain 3 g of the bispirocyclic product (yield 83%, ee 98%, dr value > 20:1).

[0072] Characterization data for 3g of product: Light yellow oil; 83% yield; >20:1 dr, 98% ee; [α] D 25=12.1(c 0.25,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3,i-PrOH / hexane=20 / 80,flow rate 1.0mL / min,λ=214nm,major diastereomer:t minor =12.4min,t major =8.3min). 1 H NMR (400MHz, CDCl3): δ7.36-7.30(m,3H),7.29-7.26(m,2H),7.14-7.10(m,2H),6.95(t,J=7.6Hz,1H),5.31-5.29(m,1H),5.01(s ,2H),4.28-4.20(m,1H),3.60(s,3H),3.51-3.45(m,1H),3.39(s,3H),2.93(q,J=7.2Hz,1H),2.54(s,3H),2.45(d,J=8.4Hz,1H). 13 C NMR (101MHz, CDCl3): δ176.1,169.6,155.4,142.7,135.7,134.9,128.7,128.6,128.5,125.9,123.0,121.7,121 .0,72.1,70.1,67.5,58.1(q,J=31.3Hz,1C),53.0,33.6,29.9(d,J=11.1Hz,1C),19.2.HRMS(ESI-TOF)calcd.for C 24 H 25 F3N3O5[M+H] + 492.1741; found: 492.1746.

[0073] Table 1: Data for 3g single crystals of the compound

[0074]

[0075] Example 8:

[0076]

[0077] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 7-fluoroindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 11 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3h (yield 96%, ee 94%, dr value >20:1).

[0078] Product characterization data over 3 hours: Light yellow oil; 96% yield; >20:1 dr, 94% ee; [α] D 25 =4.0(c0.50,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3,i-PrOH / hexane=20 / 80,flow rate 1.0mL / min,λ=254nm,major diastereomer:t minor =6.6min,t major =5.5min). 1 H NMR (400MHz, CDCl3): δ7.35-7.32(m,3H),7.28-7.25(m,2H),7.13-7.07(m,2H),6.97-6.94(m,1H),5.42(s,1H),5.00(dd,J=19.2, 12.4Hz,2H),4.30-4.22(m,1H),3.59(s,3H),3.50-3.44(m,1H),3.34(d,J=2.8Hz,3H),2.88(q,J=7.2Hz,1H)2.49(d,J=8.0Hz,1H); 13 C NMR (101MHz, CDCl3): δ175.3,169.2,155.2,149.3,146.8,135.6,131.8(d,J =9.1Hz,1C),128.6(d,J=11.1Hz,1C),128.6,128.3(d,J=4.0Hz,1C),123.5(d ,J=6.1Hz,1C),119.9(d,J=4.0Hz,1C),119.2,119.0,72.6,70.6,67.6,58.0( q,J=32.3Hz,1C),53.1,33.6,29.0(d,J=6.1Hz,C).HRMS(ESI-TOF)calcd.for C23 H 22 F4N3O5[M+H] + 496.1490; found: 496.1492.

[0079] Example 9:

[0080]

[0081] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 7-chloroindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 20 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3i (yield 99%, ee 99%, dr value > 20:1).

[0082] Product 3i characterization data: Light yellow oil; 99% yield; >20:1 dr, 99% ee; [α] D 25 =80.4(c 0.34,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3, i-PrOH / hexane=20 / 80, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =7.3min,t major =5.8min). 1 H NMR (400MHz, CDCl3): δ7.40-7.33(m,3H),7.30-7.26(m,3H),7.18(d,J=7.2Hz,1H),6.91(t,J=8.0Hz,1H),5.38(s,1H),5.00(dd,J =20.4,12.0Hz,2H),4.29-4.25(m,1H),3.60(s,3H),3.51(s,3H),3.48-.3.44(m,1H),2.87(q,J=7.2Hz,1H),2.47(d,J=8.0Hz,1H). 13C NMR (101MHz, CDCl3): δ176.0,169.1,155.2,141.0,135.6,133.3,128.7,128.7,128.6,128.2,123.5,12 2.5,116.7,72.0,70.7,67.6,58.0(q,J=32.3Hz,1C),53.1,33.7,29.9,14.3.HRMS(ESI-TOF)calcd.for C 23 H 22 ClF3N3O5[M+H] + 512.1195; found: 512.1203.

[0083] Example 10:

[0084]

[0085] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol 5,7-dimethylindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 36 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3j (yield 60%, ee 97%, dr value > 20:1).

[0086] Product 3j characterization data: Light yellow oil; 60% yield; >20:1 dr, 97% ee; [α] D 25 =6.8(c0.49,CH2Cl2).The ee was determined by HPLC(Chiralpak OZ-3, i-PrOH / hexane=20 / 80, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =11.5min,t major =7.9min). 1H NMR (400MHz, CDCl3): δ7.35-7.26(m,5H),6.94(d,J=9.6Hz,2H),5.30(s,1H),5.03(dd,J=20.8,12Hz,2H),4.27-4.19(m, 1H),3.60(s,3H),3.51-3.45(m,1H),3.36(s,3H),2.93(q,J=7.2Hz,1H),2.49(s,3H),2.44(d,J=8.4Hz,1H),2.28(s,3H); 13 C NMR (101MHz, CDCl3): δ176.0,169.7,155.4,140.2,135.8,135.4,132.5,128.7,128.5,128.4,126.0,122.4,1 20.8,72.3,70.0,67.4,58.2(dd,J=2.3Hz,1C),53.0,33.6,29.9,21.0,19.1,14.2.HRMS(ESI-TOF)calcd.for C 25 H 27 F3N3O5[M+H] + 506.1897; found: 506.1899.

[0087] Example 11:

[0088]

[0089] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol N-benzylindigo-derived trifluoromethylimine ylide, and 0.15 mmol 2-amidoacrylate were dissolved in 1 mL toluene. The mixture was stirred at 10 °C under a closed system for 36 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3K (yield 96%, ee 97%, dr value > 20:1).

[0090] Product 3K characterization data: Light yellow oil; 96% yield; >20:1 dr, 97% ee; [α] D 25=82.4(c 0.50,CH2Cl2).The ee was determined by HPLC(Chiralpak OX-3, i-PrOH / hexane=30 / 70, flow rate 1.0mL / min, λ=254nm, major diastereomer:t minor =7.8min,t major =21.1min). 1 H NMR (400MHz, CDCl3): δ7.36-7.26(m,12H),7.07(t,J=7.6Hz,1H),6.76(d,J=7.6Hz,1H),5.34(s,1H),5.07-5.01(m,3H), 4.57(d,J=15.6Hz,1H),4.33-4.22(m,1H),3.57-3.51(m,1H),3.41(s,3H),3.03(q,J=7.2Hz,1H),2.50(d,J=8.0Hz,1H); 13 C NMR (101MHz, CDCl3): δ175.4,169.5,155.5,144.4,135.7,135.4,131.2,129.0,128.7,128.6,128.6,127.9,127.7,125 .4,124.1,123.2,110.3,72.6,70.0,67.6,58.1(q,J=64.6,32.31C),53.0,44.4,33.4,29.9.HRMS(ESI-TOF)calcd.for C 29 H 27 F3N3O5[M+H] + 554.1897; found: 554.1906.

[0091] Example 12:

[0092]

[0093] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol 2-phthalimide acrylate were dissolved in 1 mL toluene. The mixture was stirred at 30 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 3l of the bispirocyclic product (yield 93%, ee 97%, dr value 80:20).

[0094] Product 3L characterization data: Light yellow oil; 93% yield; 80:20 dr, 97% ee; The ee was determined by HPLC (Chiralpak OX-3, i-PrOH / hexane=20 / 80, flow rate 1.0 mL / min, λ=214 nm, major diastereomer:t minor =17.0min,t major =14.2min). 1 H NMR (400MHz, CDCl3): δ7.81(d,J=7.2Hz,1H),7.72-7.65(m,2H),7.38(t,J=7.6Hz,1H),7.12-7.01(m,2H),6.87(d,J=7. 6Hz,1H),4.58-4.45(m,1H),4.12(q,J=7.2Hz,0H),3.76(s,3H),3.73-3.65(m,2H),3.12(s,3H),2.75(d,J=9.6Hz,1H).

[0095] Example 13:

[0096]

[0097] In a 10 mL dry reaction flask, 0.005 mmol Cu(OTf)₂, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol 2-phthalimide acrylate were dissolved in 1 mL toluene. The mixture was stirred and reacted at 30 °C under a closed system for 21 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 31 spirocyclic product (yield 65%, ee 92%, dr value 79:21).

[0098] Example 14:

[0099]

[0100] In a 10 mL dry reaction flask, 0.005 mmol CuBr, 0.0075 mmol chiral ligand L3, 0.01 mmol Cs₂CO₃, 0.1 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol 2-phthalimide acrylate were dissolved in 1 mL toluene. The mixture was stirred at 30 °C under a closed system for 58 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 3l of the bispirocyclic product (yield 63%, ee 94%, dr value 76:24).

[0101] Example 15:

[0102]

[0103] In a 10 mL dry reaction flask, 0.005 mmol AgOAc, 0.0075 mmol chiral ligand L1, 0.01 mmol Cs₂CO₃, 0.1 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol 2-phthalimide acrylate were dissolved in 1 mL toluene. The mixture was stirred and reacted at 30 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 3l of the bispirocyclic product (yield 82%, ee 16%, dr value >20:1).

[0104] Example 16:

[0105]

[0106] In a 10 mL dry reaction flask, 0.005 mmol AgOAc, 0.0075 mmol chiral ligand L9, 0.01 mmol Cs₂CO₃, 0.1 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol 2-phthalimide acrylate were dissolved in 1 mL toluene. The mixture was stirred at 30 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 31 spirocyclic product (yield 86%, ee 36%, dr value >20:1).

[0107] Example 17:

[0108]

[0109] In a 10 mL dry reaction flask, add 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, and 0.01 mmol... tBuOK, 0.1 mmol of N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol of 2-phthalimide acrylate were dissolved in 1 mL of toluene. The mixture was stirred at 30 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 3 μL of the bispirocyclic product (yield 67%, ee 94%, dr value 79:21).

[0110] Example 18:

[0111]

[0112] In a 10 mL dry reaction flask, 0.005 mmol CuCl, 0.0075 mmol chiral ligand L3, 0.01 mmol Na2CO3, 0.1 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.15 mmol 2-phthalimide acrylate were dissolved in 1 mL toluene. The mixture was stirred at 30 °C under a closed system for 48 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to give 31 μL of the bispirocyclic product (yield 20%, ee 95%, dr value 72:28).

[0113] Example 19:

[0114]

[0115] In a 10 mL dry reaction flask, 0.01 mmol CuCl, 0.015 mmol chiral ligand L3, 0.02 mmol Cs₂CO₃, 0.2 mmol N-methylindigo-derived trifluoromethylimide ylide, and 0.3 mmol 2-phthalimide acrylate were dissolved in 2 mL toluene. The mixture was stirred at 30 °C under a closed system for 36 hours, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the mixture was purified by column chromatography to obtain the bispirocyclic product 3M (yield 54%, ee 93%, dr value 55:45).

[0116] Antitumor activity verification:

[0117] Compound 3d from Example 4 was selected as a representative, and its in vitro antitumor activity was tested using human hepatocellular carcinoma cell line (HepG2), human breast cancer cells (MDA-MB-231), and human cervical cancer cells (HeLa) as receptors (MTT colorimetric method).

[0118] The following explanation uses compound 3d from Example 4 as an example. The specific experimental procedures are as follows:

[0119] (1) Human hepatocellular carcinoma cell line (HepG2), human breast cancer cells (MDA-MB-231), and human cervical cancer cells (HeLa) in logarithmic growth phase were used to adjust the cell density to 1×10⁻⁶ cells / year using fresh DMEM medium containing 10% fetal bovine serum. 5 cells·mL -1 Inoculate into 96-well plates at 100 μL / well and incubate at 37°C in a 5% CO2 incubator.

[0120] (2) After 24 hours of adherent cell culture, the medium was replaced with serum-free fresh medium, and drugs were added simultaneously. The drug dosages for the samples were 0.4 μmol / mL, 2 μmol / mL, 10 μmol / mL and 50 μmol / mL, and a positive control group was also included.

[0121] (3) After the cells were drugged, they were cultured for 48 hours. Then, MTT staining solution (10 μL, 0.5 mg / mL) was added to the culture medium and the cells were cultured at 37°C for another 4 hours.

[0122] (4) Remove the culture medium, add 100 μL of dimethyl sulfoxide to dissolve the formalin crystals, measure the OD value at 570 nm using a microplate reader (ELx800, Bio-Tek, Winooski, VT, USA), and calculate the IC50 of compound 4 at 3 days. 50 value.

[0123] The tumor cell inhibitory activity experiments of the compounds in other examples were conducted in a similar manner to those in Example 4.

[0124] Table 1 shows the inhibitory effects of some of the compounds in the examples on the proliferation of human liver cancer cell line (HepG2), human breast cancer cells (MDA-MB-231), and human cervical cancer cells (HeLa).

[0125]

[0126] The above experiments show that the compounds involved in this invention have a certain degree of inhibitory effect on human liver cancer cells, human breast cancer cells, and human cervical cancer cells.

[0127] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific structures and characteristics is not elaborated upon here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the structure 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 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 spiro[indole-pyrrolidine amino acid ester] compound containing trifluoromethyl, characterized in that: The general structural formula of the compound is: ； In the formula: Ar 1 It is selected from any one of 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 5-fluorophenyl, 5-chlorophenyl, 5-bromophenyl, 6-fluorophenyl, 6-chlorophenyl, 6-bromophenyl, 7-fluorophenyl, 7-chlorophenyl, and 7-bromophenyl; R 1 Selected from methyl or benzyl; R 2 R 3 All are selected from either hydrogen or benzyloxycarbonyl, and R 2 R 3 They are not the same; R is a methyl group.

2. The method for preparing the chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl according to claim 1, characterized in that... Includes the following steps: Step 1: Add CuCl catalyst, chiral ligand L3, cesium carbonate, indigo-derived trifluoromethylimine ylide and 2-amidoacrylate to the reaction tube in sequence, then add toluene. Stir the mixture at 10°C. Step 2: After the reaction is complete, the chiral spiro[oxyindole-pyrrolidine amino acid ester] compound containing trifluoromethyl is obtained by post-processing and separation. The synthesis route is shown below: ； Among them, Ar 1 , R, R 1 R 2 R 3 The choice is as described in claim 1; The structure of the chiral ligand L3 is as follows: .

3. The application of the trifluoromethyl-containing chiral spiro[oxyindole-pyrrolidine amino acid ester] compound according to claim 1 in the preparation of antitumor drugs, wherein the tumor is liver cancer, breast cancer, or cervical cancer.

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