Trifluoromethyl cyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative as well as preparation method and application thereof

Abstract

The invention discloses a trifluoromethyl cyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative as well as a preparation method and application thereof, and belongs to the technical field of organic compound synthesis and medicine. The synthesis method of the trifluoromethyl cyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative comprises the following two steps: firstly, carrying out a Mitsunobu reaction on a benzyl alcohol compound and hydroxy naphthaldehyde (or hydroxy benzaldehyde), and then carrying out a reaction on the obtained product and amino-acid ester hydrochloride (or sulfamic acid) to obtain the trifluoromethyl cyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative. The Schiff base is reduced with sodium cyanoborohydride (or simple hydrolysis is carried out after reduction), and the Schiff base is obtained. The preparation method provided by the invention has the characteristics of simple steps, mild reaction conditions and fast reaction, and meets the requirements of green chemistry. The compound prepared according to the method can selectively regulate the S1P1 receptor without exciting the S1P3 receptor, and is expected to be developed into a novel preparation for treating inflammatory immune related diseases such as pulmonary fibrosis, ulcerative colitis and multiple sclerosis.

Description

Technical Field

[0001] The invention belongs to the technical field of organic compound synthesis and medicine, and specifically relates to a nitrogen-containing organic acid derivative substituted with trifluoromethylcyclohexylbenzyl ether, a synthesis method thereof and pharmaceutical use thereof. Background Art

[0002] Sphingosine-1 phosphate (S1P) is a member of the family of five G protein-coupled receptors on the cell membrane. 1-5 ), which can regulate a variety of biological processes, including cell differentiation, vascular stability, inflammation, endothelial integrity and angiogenesis. Sphingosine-1-phosphate (S1P) is a biologically active sphingolipid substance that is involved in the occurrence and development of many diseases, especially immune-mediated diseases, and has become an important drug target. Studies have shown that selectively regulating S1P1 without exciting S1P3 can be a new way to treat inflammatory-related diseases such as idiopathic pulmonary fibrosis (IPF), ulcerative colitis, and multiple sclerosis.

[0003] Therefore, by exploring the structural characteristics of S1P1, simulating the binding mode of compounds with receptors, and targeting the design and synthesis of new selective S1P1 modulators, it is expected that effective drugs will be discovered for the treatment of inflammatory-related diseases such as pulmonary fibrosis, ulcerative colitis, and multiple sclerosis. Summary of the invention

[0004] In view of the above technical problems, the present invention designs a series of nitrogen-containing organic acid derivatives substituted with trifluoromethyl cyclohexyl benzyl ether. The compounds have the structural characteristics of a lipophilic group cyclohexane trifluoromethyl benzyl ether as a tail, a naphthalene ring or a benzene ring as a connecting aromatic region, and an amino acid, aminosulfonic acid or aminophosphoric acid as a hydrophilic polar head. At the same time, the compounds can selectively regulate S1P1 receptors and have high application value in the field of medicine.

[0005] The present invention also provides a method for synthesizing the nitrogen-containing organic acid derivative substituted with trifluoromethylcyclohexylbenzyl ether.

[0006] The present invention also provides a pharmaceutical use of the nitrogen-containing organic acid derivative substituted with the trifluoromethylcyclohexylbenzyl ether.

[0007] According to one aspect of the present invention, a trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative is provided, including a compound having a structure shown in formula (I) and a pharmaceutically acceptable salt thereof:

[0008]

[0009] in,

[0010] R1 is selected from one of phenyl and naphthyl;

[0011] R2 is selected from

[0012] One of;

[0013] R3 is selected from one of H and methyl.

[0014] In some embodiments of the present invention, in the formula (I), when R1 is selected from a naphthalene ring, R3 is selected from one of hydrogen or methyl or other substitutions, and R2 is selected from

[0015] One of them.

[0016] In some embodiments of the present invention, in the formula (I), when R1 is selected from a benzene ring, R3 is selected from hydrogen, and R2 is selected from One of them.

[0017] In some embodiments of the present invention, the trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivatives Ⅰ1-12, Ⅱ1-6, Ⅲ1-4, at least one of the compounds of structures shown in Ⅳ1:

[0018]

[0019]

[0020]

[0021] According to another aspect of the present invention, a method for synthesizing the trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative is provided, comprising firstly using a benzyl alcohol compound of formula (Ia) and hydroxynaphthaldehyde (or hydroxybenzaldehyde) to carry out a Mitsunobu reaction to generate formula (Ib), then reacting with an amino acid (or aminosulfonic acid, or aminophosphoric acid) to generate a Schiff base, and then reducing with sodium cyanoborohydride to obtain the trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid compound of formula (I).

[0022]

[0023] In some preferred embodiments of the present invention, the hydroxynaphthaldehyde (or hydroxybenzaldehyde) compound includes at least one of the compounds shown in the following formula:

[0024]

[0025] In some embodiments of the present invention, the reaction for synthesizing the compound of formula (Ib) is carried out in the presence of anhydrous tetrahydrofuran under the catalysis of triphenylphosphine and diisopropyl azodicarboxylate.

[0026] In some embodiments of the present invention, the reaction temperature for synthesizing formula (Ib) is room temperature (5-30°C).

[0027] In some embodiments of the present invention, the reaction of steps (2) and (3) is carried out in anhydrous dichloromethane and methanol in the presence of acetic acid, sodium cyanoborohydride and N,N-diisopropylethylamine.

[0028] In some embodiments of the present invention, the reaction temperature of steps (2) and (3) is room temperature (5-30°C).

[0029] In some embodiments of the present invention, the reaction of step (4) is carried out in an aqueous solution of lithium hydroxide in the presence of anhydrous methanol.

[0030] In some embodiments of the present invention, the reaction temperature for synthesizing formula (I) is room temperature (5-30°C).

[0031] In some embodiments of the present invention, the reaction duration is 12 h (overnight).

[0032] In some embodiments of the present invention, the reaction duration is 2h-6h.

[0033] In some embodiments of the present invention, the synthesis method further comprises purifying the obtained compound after the reaction.

[0034] In some embodiments of the present invention, the purification comprises extraction, column chromatography or PTLC separation, filtration and drying.

[0035] In some embodiments of the present invention, the extractant used in the extraction includes dichloromethane and ethyl acetate.

[0036] In some embodiments of the present invention, the column chromatography used is silica gel column chromatography.

[0037] Beneficial technical effects

[0038] 1. The present invention discloses a novel trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative, and the synthesis method of the present invention has the characteristics of simple steps, mild reaction conditions and fast reaction;

[0039] 2. The present invention experimentally studies the agonist activity of nitrogen-containing organic acid derivatives substituted with trifluoromethylcyclohexylbenzyl ether on S1P1, and proves that compounds I6, I11, I12, II3, II6, and III4 have high selectivity, strong agonist activity on S1P1, and no agonist activity on S1P3;

[0040] 3. The present invention uses trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivatives to study the efficacy of pulmonary fibrosis in mice, proving that I12 has a significant anti-fibrosis effect;

[0041] 4. The present invention studies the efficacy of nitrogen-containing organic acid derivatives substituted with trifluoromethylcyclohexylbenzyl ether on colitis in mice, proving that I12 has a significant anti-colitis effect; BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 .The effects of I12 and II3 on ​​peripheral blood lymphocytes of rats (8 hours), compared with the control group, *P<0.05, ***P<0.001;

[0043] Figure 2 .The effects of various concentrations of I12 on peripheral blood lymphocytes of rats (12 hours) were compared with those of the control group, *P<0.05, ***P<0.001;

[0044] Figure 3 .Compared with the control group, the effects of different concentrations of I12 on the dynamic compliance of mouse lungs were ####P<0.0001; compared with the model group, *P<0.05, ***P<0.001;

[0045] Figure 4 .Compared with the control group, the effect of I12 at different concentrations on the dynamic lung resistance of mice was #P<0.05; compared with the model group, *P<0.05;

[0046] Figure 5 .Compared with the control group, the effects of different concentrations of I12 on the dynamic elasticity of mice were ##P<0.01; compared with the model group, *P<0.05, **P<0.01;

[0047] Figure 6 .Compared with the control group, the effects of different concentrations of I12 on the deep inspiration volume of mice were ####P<0.0001; compared with the model group, *P<0.05, **P<0.01;

[0048] Figure 7 .Representative images of the effects of various I12 concentrations on mouse colon length;

[0049] Figure 8.Compared with the control group, the effect of different concentrations of I12 on the colon length of mice was ####P<0.0001; compared with the model group, *P<0.05;

[0050] Fig. 9 .Compared with the control group, the effects of different concentrations of I12 on the DAI score of mice were ####P<0.0001; compared with the model group, **P<0.01, ***P<0.001;

[0051] Fig.10 .Compared with the control group, the effect of each concentration of I12 on the percentage of weight loss of mice was ####P<0.0001; compared with the model group, *P<0.05, **P<0.01;

[0052] Fig.11 .Compared with the control group, the effect of different concentrations of I12 on the stool shape of mice was ####P<0.0001; compared with the model group, **P<0.01, ***P<0.001;

[0053] Fig.12 .Compared with the control group, the effects of different concentrations of I12 on the degree of bloody stool in mice were ####P<0.0001; compared with the model group, ***P<0.001. DETAILED DESCRIPTION

[0054] The present invention will be fully described below in conjunction with the embodiments to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0055] Example 1

[0056] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula Ⅰ1, and the specific process is as follows:

[0057] S1. 3-trifluoromethyl-4-cyclohexylbenzyl alcohol (5.0mmol, 1290mg, CAS: 957205-23-1), 6-hydroxy-2-naphthaldehyde (5.0mmol, 860mg, CAS: 78119-82-1) and triphenylphosphine (PPh3, 6.0mmol, 1570mg, CAS: 603-35-0) were dissolved in anhydrous tetrahydrofuran (10mL, CAS: 109-99-9), stirred at room temperature, and diisopropyl azodicarboxylate (DIAD) (6.0mmol, 1210mg, CAS: 7087-68-5) was slowly added dropwise within 5min, and stirring was continued. The reaction was allowed to react overnight at room temperature. After the reaction was completed, the reaction solvent was dried and separated by column chromatography (eluent was petroleum ether: ethyl acetate = 20:1) to obtain compound 3a, 1230mg of white powdery solid, with a yield of 60%.

[0058] S2. Compound 3a (0.2mmol, 82mg) and glycine methyl ester hydrochloride (0.3mmol, 38mg, CAS: 5680-79-5) were dissolved in methanol / dichloromethane (V:V = 2mL: 2mL), N, N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg, CAS: 7087-68-5) was added, and sodium cyanoborohydride (0.3mmol, 19mg, CAS: 25895-60-7) and acetic acid (0.8mmol, 48mg, CAS: 64-19-7) were added after stirring at room temperature for 2h, and stirring was continued. TLC was used to monitor the progress of the reaction. When 3a was consumed, saturated sodium bicarbonate solution (20mL, CAS: 144-55-8) was added, and then extracted with ethyl acetate, the organic phase was collected, concentrated under reduced pressure, and separated by PTLC (developing solvent was petroleum ether: ethyl acetate = 3:1) to obtain compound 4a as a colorless oily solid.

[0059] S3. Dissolve compound 4a obtained in S2 in methanol (4 mL, CAS: 67-56-1), add aqueous lithium hydroxide solution (0.5 mol / L, 3.2 mL, CAS: 1310-65-2), and stir at room temperature for 3 h. After the reaction, add 2 mol / L dilute hydrochloric acid (CAS: 7647-01-0) to adjust the pH to 1-2, continue to add water until a precipitate is formed, let stand for 10 h, and filter to obtain compound Ⅰ1, 60 mg of white powder, with a yield of 64%.

[0060] The reaction that occurs in this embodiment is shown in the following formula:

[0061]

[0062] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula Ⅰ1 is indeed obtained in this example.

[0063] NMR data of compound 3a, 1 H NMR (600 MHz, CDCl3-d 1 )δ1.25-1.33(m,1H),1.39-1.49(m,4H),1.77(d,J=12.6Hz,1H),1.82-1.86(m,4H),2.93-2.97(m,1H),7.26(d,J=2.4Hz,1H),7.30(dd,J=2.4 ,9.0Hz,1H),7.50(d,J=8.4Hz,1H),7.60(d,J=7.8Hz,1H),7.72(s,1H),7.80(d,J=8.4Hz,1H),7.91-7.93(m,2H),8.26(s,1H),10.10(s,1H); 13 C NMR (150MHz, CDCl3-d 1 )δ26.08,26.79,34.54,40.01,40.02,69.46,107.35,120.12,123.76,124.92,124.96,125.00,125.04,127.86,128.19,128.57,131.02,131.33,132.57,133.76,134.22,138.16,147.21,159.14. High resolution mass spectrometry data: HRMS (ESI) calcd for C 25 H 23 F3O2(M+H) + :413.1723,found413.1728.

[0064] NMR data of compound Ⅰ1, 1 H NMR (600 MHz, MeOD-d 4)δ1.33-1.45(m,3H),1.51-1.57(m,2H),1.77(d,J=12.0Hz,3H),1.85-1.87(m, 2H),2.91(t,J=11.4Hz,2H),3.94(s,2H),4.39(s,2H),5.24(s,2H),7.29(dd,J= 2.4,9.0Hz,1H),7.39(d,J=2.4Hz,1H),7.53(dd,J=0.6,8.4Hz,1H),7.57(d,J= 8.4Hz,1H),7.68(d,J=8.4Hz,1H),7.46(s,1H),7.85-7.88(m,2H),7.94(s,1H); 13 C NMR (150 MHz, MeOD-d 4 )δ27.12,28.02,35.63,41.53,47.71,48.62,48.76,48.90,49.05,49.19,49. 33,49.47,52.30,70.21,108.34,121.03,125.28,125.63,125.67,125.71,125 .75,127.09,127.25,128.32,128.71,128.90,129.25,129.77,130.32,130.92,130.93,132.48,136.51,147.86,159.03,168.89. High resolution mass spectrometry data: HRMS (ESI) calcd for C 27 H 28 F3NO3(M+H) + :472.2094,found472.2097.

[0065] Example 2

[0066] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I2, and the specific process is as follows:

[0067] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0068] S2. Compound 3a (0.2mmol, 82mg) and aspartic acid methyl ester hydrochloride (0.3mmol, 60mg, CAS: 69630-50-8) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4b as a colorless oily solid.

[0069] S3. Compound 4b obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ2, 27 mg of white powder, with a yield of 26%.

[0070] The reaction that occurs in this embodiment is shown in the following formula:

[0071]

[0072] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula Ⅰ2 is indeed obtained in this example.

[0073] NMR data of compound Ⅰ2, 1 H NMR (600 MHz, DMSO-d 6 )δ1.30-1.35(m,3H),1.52-1.56(m,2H),1.69(d,J=11.4Hz,3H),1.80(d,J=12.0Hz,2H),2.60-2 .64(m,1H),2.72(dd,J=9.0,16.2Hz,1H),2.80(t,J=11.4Hz,1H),3.68-3.70(m,1H),4.10(d,J= 13.2Hz,1H),4.19(d,J=13.2Hz,1H),5.27(s,2H),7.26-7.28(m,1H),7.45(d,J=1.8Hz,1H),7.5 2(d,J=8.4Hz,1H),7.65(d,J=8.4Hz,1H),7.75(d,J=8.4Hz,1H),7.79(s,1H),7.78-7.87(m,3H); 13 C NMR (150 MHz, DMSO-d 6)δ25.85,26.88,34.31,36.73,39.56,39.70,39.84,39.97,40.11,40.25,40.39,5 0.14,56.03,68.85,107.65,119.60,124.18,125.17,125.21,125.25,125.29,126 .00,126.62,126.81,127.51,128.06,128.63,128.72,129.19,129.95,130.62,132.55,134.33,135.61,146.40,156.92,171.57,171.72. High resolution mass spectrometry data: HRMS (ESI) calculation for C 29 H 30 F3NO5(M+H) + :530.2149, found 530.2158.

[0074] Example 3

[0075] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I3, and the specific process is as follows:

[0076] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0077] S2. Compound 3a (0.2mmol, 82mg) and leucine methyl ester hydrochloride (0.3mmol, 60mg, CAS: 7517-19-3) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4c as a colorless oily solid.

[0078] S3. Compound 4c obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL, CAS: 1310-65-2) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ3 as a white powder (89 mg). The yield was 85%.

[0079] The reaction that occurs in this embodiment is shown in the following formula:

[0080]

[0081] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula Ⅰ3 is indeed obtained in this example.

[0082] NMR data of compound Ⅰ3, 1 H NMR (600 MHz, MeOD-d 4 )δ0.92(d,J=6.0Hz,1H),0.97(d,J=6.0Hz,1H),1.34-1.57(m,5H),1.66-1.71(m,1 H),1.77-1.87(m,7H),2.91(t,J=11.6Hz,1H),3.75-3.78(m,1H),4.28-4.35(m,2H ),5.22(s,3H),7.27(d,J=9.0Hz,1H),7.37(s,1H),7.52(d,J=8.4Hz,1H),7.56(d, J=8.4Hz,1H),7.67(d,J=7.8Hz,1H),7.73(s,1H),7.83-7.86(m,2H),7.91(s,1H); 13 C NMR (150 MHz, MeOD-d 4 )δ20.77,21.75,24.63,25.68,26.57,34.19,39.08,40.08,50.20,59.30,68.75,106.88,119.52,123.84,124.18,124.22,124.26,124.30,125.65,126.14,127.03,127.26,127.45,127.67,128.31,128.85,129.43,129.55,131.02,134.99,135.07,146.41,157.53. High resolution mass spectrometry data: HRMS (ESI) calcd for C 31 H 36 F3NO3(M+H) + :528.272, found 528.2734.

[0083] Example 4

[0084] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I4, and the specific process is as follows:

[0085] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0086] S2. Compound 3a (0.2mmol, 82mg) and proline methyl ester hydrochloride (0.3mmol, 50mg, CAS: 2133-40-6) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4d as a colorless oily solid.

[0087] S3. Compound 4d obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ4 as a yellow solid (57 mg). The yield was 56%.

[0088] The reaction that occurs in this embodiment is shown in the following formula:

[0089]

[0090] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula Ⅰ4 is indeed obtained in this example.

[0091] NMR data of compound Ⅰ4, 1 H NMR (600 MHz, MeOD-d 4 )δ1.32-1.55(m,5H),1.78-1.88(m,5H),1.92-1.97(m,1H),2.10-2.17(m,2H),2.45-2.51 (m,1H),2.92-2.96(m,1H),3.25-3.30(m,1H),3.54-3.58(m,1H),3.96-3.98(m,1H),4.38( d,J=12.6Hz,1H),4.58(d,J=12.6Hz,1H),5.22(s,2H),7.27(d,J=8.4Hz,1H),7.36(s,1H), 7.57-7.58(m,2H),7.67(d,J=7.2Hz,1H),7.75(s,1H),7.83(d,J=7.8Hz,2H),7.96(s,1H); 13 C NMR (150 MHz, MeOD-d 4)δ12.00,12.44,23.68,25.70,38.19,40.85,41.02,53.44,65.27,82.19,112.74,122.59,123.13,127.07,131.25,131.27,132.47,133.30,139.74,146.25,151.56,163.87,164.07,178.03. High resolution mass spectrometry data: HRMS (ESI) calcd for C 29 H 32 F3NO4(M+H) + :512.2407,found512.5037.

[0092] Example 5

[0093] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I5, and the specific process is as follows:

[0094] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0095] S2. Compound 3a (0.2mmol, 82mg) and sarcosine methyl ester hydrochloride (0.3mmol, 42mg, CAS: 13515-93-0) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4e as a colorless oily solid.

[0096] S3. Compound 4e obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ5 as a white oil (81 mg). The yield was 84%.

[0097] The reaction that occurs in this embodiment is shown in the following formula:

[0098]

[0099] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula I5 is indeed obtained in this example.

[0100] NMR data of compound Ⅰ5, 1 H NMR (600 MHz, MeOD-d 4 )δ1.34-1.58(m,5H),1.77-1.88(m,5H),2.93(s,3H),3.31(s,2H),4.10(s,2H),4.55(s,1H),5.26(s,2H),7.31(dd,J=2.4,9 .0Hz,1H),7.41(d,J=2.4Hz,1H),7.57-7.59(m,2H),7.69(d,J=8.4Hz,1H),7.75(s,1H),7.88(t,J=9.0Hz,1H),8.00(s,3H); 13 C NMR (150 MHz, MeOD-d 4 )δ7.12,28.02,35.62,41.44,41.53,56.38,61.62,70.22,108.35,121.16,125.28,125.62,125.67,125.71,125.74,127.09,128.70,128.90,129.16,129.30,129.79,130.23,131.07,132.46,132.50,136.49,136.79,147.87,159.24,168.49. High resolution mass spectrometry data: HRMS (ESI) calcd for C 28 H 30 F3NO3(M+H) + :486.2251,found486.2259.

[0101] Example 6

[0102] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I6, and the specific process is as follows:

[0103] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0104] S2. Compound 3a (0.2mmol, 82mg) and dimethyl glutamate hydrochloride (0.3mmol, 64mg, CAS: 23150-65-4) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4f as a colorless oily solid.

[0105] S3. Dissolve compound 4f obtained in S2 in methanol (4 mL), add aqueous lithium hydroxide solution (0.5 mol / L, 3.2 mL), and stir at room temperature for 3 h. After the reaction, add 2 mol / L dilute hydrochloric acid to adjust the pH to 1-2. Continue to add water until a precipitate is formed. Let stand for 10 h, and filter to obtain compound Ⅰ6 as a yellow solid, 54 mg, with a yield of 50%.

[0106] The reaction that occurs in this embodiment is shown in the following formula:

[0107]

[0108] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula Ⅰ6 is indeed obtained in this example.

[0109] NMR data of compound Ⅰ6, 1 H NMR (600 MHz, MeOD-d 4 )δ1.34-1.57(m,5H),1.77-1.88(m,5H),2.06-2.11(m,1H),2.30-2.35(m,1H),2.42-2.4 7(m,1H),2.52-2.58(m,1H),2.91(t,J=11.4Hz,1H),4.0(dd,J=3.0,9.6Hz,1H),4.12(d, J=15.0Hz,1H),5.10(d,J=15.0Hz,1H),5.21(s,2H),7.21(dd,J=2.4,9.0Hz,1H),7.29-7 .32(m,2H),7.56(d,J=8.4Hz,1H),7.63(s,1H),7.67(d,J=7.8Hz,1H),7.74-7.76(m,3H); 13 C NMR (150 MHz, MeOD-d 4)δ24.01,27.15,28.04,30.76,35.64,41.53,46.76,60.58,70.15,108.33,120.36,125.31,125.62,125.66,125.70,125.74,127.12,127.78,128.33,128.79,129.74,130.50,130.53,132.41,132.46,135.66,136.72,147.79,158.27,175.10,178.12. High resolution mass spectrometry data: HRMS (ESI) calcd for C 30 H 32 F3NO5(M-H2O+H) + :526.2200, found 526.2208.

[0110] Example 7

[0111] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I7, and the specific process is as follows:

[0112] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0113] S2. Compound 3a (0.2mmol, 82mg) and phenylalanine methyl ester hydrochloride (0.3mmol, 65mg, CAS: 7524-50-7) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), and N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added. After stirring at room temperature for 2h, sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4g as a colorless oily solid.

[0114] S3. Dissolve 4 g of compound obtained in S2 in methanol (4 mL), add aqueous lithium hydroxide solution (0.5 mol / L, 3.2 mL), stir at room temperature for 3 h, add 2 mol / L dilute hydrochloric acid after the reaction to adjust the pH to 1-2, continue to add water until a precipitate is formed, let stand for 10 h, and filter to obtain compound Ⅰ7, 100 mg of white powder, with a yield of 90%.

[0115] The reaction that occurs in this embodiment is shown in the following formula:

[0116]

[0117] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula Ⅰ7 is indeed obtained in this example.

[0118] NMR data of compound Ⅰ7, 1 H NMR (600 MHz, MeOD-d 4 )δ1.77-1.85(m,5H),2.92-2.95(m,1H),3.28-3.32(m,1H),3.55(d,J=5.4,1 H),3.84-3.86(m,1H),4.13(d,J=13.2Hz,1H),4.23(m,1H),4.95(s,2H),6.9 9(d,J=2.4Hz),7.12-7.14(m,3H),7.21-7.23(m,3H),7.47(d,J=7.8Hz,1H), 7.53(d,J=7.8Hz,1H),7.66-7.68(m,3H),7.70(d,J=9.0Hz,1H),7.78(S,1H); 13 C NMR (150 MHz, MeOD-d 4 )δ26.08,26.79,34.54,36.04,39.98,39.99,50.30,52.74,60.00,69.14,106.67 ,119.68,121.81,123.63,124.44,124.81,124.84,124.88,124.92,125.45,127.5 4,127.86,127.93,128.15,128.45,128.52,128.75,129.34,129.84,130.65,130.93,134.03,134.19,134.75,147.01,157.38,168.19. HRMS (ESI) calcd for C 34 H 34 F3NO3(M+H) + :562.2564, found 562.2576.

[0119] Example 8

[0120] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I8, and the specific process is as follows:

[0121] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0122] S2. Compound 3a (0.2mmol, 82mg) and isoleucine methyl ester hydrochloride (0.3mmol, 55mg, CAS: 18598-74-8) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4h as a colorless oily solid.

[0123] S3. Dissolve compound 4h obtained in S2 in methanol (4mL), add aqueous lithium hydroxide solution (0.5mol / L, 3.2mL), and stir at room temperature for 3h. After the reaction, add 2mol / L dilute hydrochloric acid to adjust the pH to 1-2. Continue to add water until a precipitate is formed. Let stand for 10h, and filter to obtain compound Ⅰ8, 39mg of white powder, with a yield of 38%.

[0124] The reaction that occurs in this embodiment is shown in the following formula:

[0125]

[0126] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula Ⅰ8 is indeed obtained in this example.

[0127] NMR data of compound Ⅰ8, 1 H NMR (600 MHz, MeOD-d 4 )δ0.94-0.98(m,6H),1.37-1.57(m,5H),1.77-1.88(m,5H),2.07-2.08(m ,1H),2.91(t,J=11.4Hz,1H),3.72(s,2H),4.05(d,J=3.6Hz,1H),4.39(s, 2H),5.24(s,2H),7.29-7.31(m,1H),7.39(d,J=2.4Hz,1H),7.53-7.59(m ,2H),7.68(d,J=7.8Hz,1H),7.74(s,1H),7.85-7.87(m,2H),7.94(s,1H); 13 C NMR (150 MHz, MeOD-d 4)δ12.05,14.53,27.13,27.78,28.02,35.64,37.63,41.53,52.57,53.47,64.65 ,65.41,70.23,108.35,120.00,121.07,125.29,125.64,125.68,125.72,125.76 ,126.66,127.10,127.13,128.16,128.78,129.13,129.78,130.22,130.91,131.62,132.50,136.50,136.56,147.87,159.12,169.34. High resolution mass spectrometry data: HRMS (ESI) calculation for C 30 H 34 F3NO3(M+H) + :514.2564,found 514.2568.

[0128] Example 9

[0129] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I9, and the specific process is as follows:

[0130] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0131] S2. Compound 3a (0.2mmol, 82mg) and tryptophan methyl ester hydrochloride (0.3mmol, 76mg, CAS: 14907-27-8) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4i as a colorless oily solid.

[0132] S3. Compound 4i obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ9 as a yellow solid (33 mg). The yield was 28%.

[0133] The reaction that occurs in this embodiment is shown in the following formula:

[0134]

[0135] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula Ⅰ9 is indeed obtained in this example.

[0136] NMR data of compound Ⅰ9, 1 H NMR (600 MHz, MeOD-d 4 )δ1.28-1.56(m,5H),1.69-1.82(m,5H),2.81(t,J=11.4Hz,1H),3.25(s ,2H),3.71(s,1H),4.06(s,2H),5.26(s,2H),6.92-6.94(m,1H),7.04-7. 07(m,1H),7.23-7.26(m,2H),7.34(d,J=7.8Hz,1H),7.44(s,1H),7.51( d,J=7.8Hz,2H),7.65(d,J=8.4Hz,1H),7.73-7.79(m,5H),10.98(s,1H); 13 C NMR (150 MHz, MeOD-d 4 )δ25.28,26.31,33.74,49.85,60.05,68.26,107.06,111.30,118.25,118.29,118.93,120.85,123.61,124.18,124.62,124.66,125.43126.04,126.23,126.78,127.07,127.59,128.06,128.61,129.32,131.96,133.71,135.05,136.05,145.81,156.30. High resolution mass spectrometry data: HRMS (ESI) calcd for C 36 H 35 F3N2O3(M+H) + :601.2673,found601.2681.

[0137] Example 10

[0138] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I10, and the specific process is as follows:

[0139] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0140] S2. Compound 3a (0.2mmol, 82mg) and tyrosine methyl ester hydrochloride (0.3mmol, 70mg, CAS: 3417-91-2) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), and N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added. After stirring at room temperature for 2h, sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4j as a colorless oily solid.

[0141] S3. Compound 4j obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ10 as a yellow solid (52 mg). The yield was 45%.

[0142] The reaction that occurs in this embodiment is shown in the following formula:

[0143]

[0144] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula Ⅰ10 is indeed obtained in this example.

[0145] NMR data of compound Ⅰ10, 1 H NMR (600 MHz, MeOD-d 4 )δ1.33-1.58(m,5H),1.75-1.87(m,5H),2.89-2.93(m,1H),3.11-3.21(m,1H),3 .98(t,J=6.6Hz,1H),4.29(d,J=2.4Hz,2H),5.26(s,2H),6.74(d,J=8.4Hz,2H),7 .09(d,J=8.4Hz,2H),7.30(dd,J=2.4,9.0Hz,1H),7.44(1s,1H),7.51(d,J=8.4Hz ,1H),7.61(d,7.8Hz,1H),7.72(d,J=7.8Hz,1H),7.77(s,1H),7.84-7.87(m,3H); 13 C NMR (150 MHz, MeOD-d 4)δ26.60,27.54,35.12,35.76,41.00,51.08,62.11,69.69,108.05,116.36 ,120.55,124.83,125.37,125.42,125.45,125.49,126.13,126.65,127.34, 127.94,128.13,128.51,129.52,129.59,130.59,130.70,131.39,132.45,135.79,136.11,147.35,157.68,158.35,170.95. High resolution mass spectrometry data: HRMS (ESI) calcd for C 34 H 34 F3NO4(M+H) + :578.2513, found 578.2527.

[0146] Embodiment 11

[0147] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I11, and the specific process is as follows:

[0148] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0149] S2. Compound 3a (0.2mmol, 82mg) and methionine methyl ester hydrochloride (0.3mmol, 60mg, CAS: 2491-18-1) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 4k as a colorless oily solid.

[0150] S3. Compound 4k obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ11 as a white powder (88 mg). The yield was 81%.

[0151] The reaction that occurs in this embodiment is shown in the following formula:

[0152]

[0153] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula Ⅰ11 is indeed obtained in this example.

[0154] NMR data of compound Ⅰ11, 1 H NMR (600 MHz, MeOD-d 4 )δ1.34-1.57(m,5H),1.77-1.87(m,5H),2.08(s,3H),2.24-2.60(m,2H),2. 60-2.71(m,2H),2.91-2.95(m,1H),4.14-4.16(m,1H),4.36-4.43(m,2H),5 .24(s,2H),7.29(dd,J=2.4,9.0Hz,1H),7.38(d,J=1.8Hz,1H),7.55-7.58( m,2H),7.68(d,J=8.4Hz,1H),7.74(s,1H),7.84-7.88(m,2H),7.95(s,1H); 13 CNMR (150MHz, MeOD-d 4 )δ13.68,25.68,26.57,28.89,29.06,34.19,40.08,50.47,58.32,68.76,106.90,119.57,123.83,124.18,124.22,124.26,124.30,125.65,125.82,127.05,127.26,127.45,127.74,128.33,128.86,129.47,129.68,131.04,135.06,135.08,146.41,157.59,169.65. High resolution mass spectrometry data: HRMS (ESI) calcd for C 30 H 34 F3NO3S(M+H) + :546.2284, found 546.2296.

[0155] Example 12

[0156] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula I12, and the specific process is as follows:

[0157] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0158] S2. Compound 3a (0.2mmol, 82mg) and threonine methyl ester hydrochloride (0.3mmol, 51mg, CAS: 62076-66-8) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The reaction progress was monitored by TLC, and the purification method was the same as that of Example 1-S2 to obtain compound 41 as a colorless oily solid.

[0159] S3. The compound 41 obtained in S2 was dissolved in methanol (4 mL, CAS: 67-56-1), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound Ⅰ12 as a yellow solid (45 mg). The yield was 44%.

[0160] The reaction that occurs in this embodiment is shown in the following formula:

[0161]

[0162] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula Ⅰ12 is indeed obtained in this example.

[0163] NMR data of compound Ⅰ12, 1 H NMR (600 MHz, MeOD-d 4 )δ1.29(d,J=6.0Hz,3H),1.36-1.57(m,5H),1.77-1.87(m,5H),2.91(t,J=11.4Hz, 1H),3.53(d,J=7.2Hz,1H),4.08-4.10(m,1H),4.36-4.44(m,2H),5.23(s,2H),7.28 (dd,J=2.4,9Hz,1H),7.37(d,J=2.4Hz,1H),7.52(dd,J=1.2,8.4Hz,1H),7.57(d,J= 8.4Hz,1H),7.68(d,J=8.4Hz,1H),7.44(s,1H),7.83(t,J=9.0Hz,2H),7.91(s,1H); 13 C NMR (150 MHz, MeOD-d 4)δ26.08,26.79,34.54,36.04,39.98,39.99,50.30,52.74,60.00,69.14,106.67 ,119.68,121.81,123.63,124.44,124.81,124.84,124.88,124.92,125.45,127.5 4,127.86,127.93,128.15,128.45,128.52,128.75,129.34,129.84,130.65,130.93,134.03,134.19,134.75,147.01,157.38,168.19. HRMS (ESI) calcd for C 29 H 32 F3NO4(M+H) + :516.2356, found 516.2364.

[0164] Example 13

[0165] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminosulfonic acid compound, the specific structure of which is shown in Formula II1, and the specific process is as follows:

[0166] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0167] S2. Dissolve compound 3a (0.2mmol, 82mg) and aminomethanesulfonic acid (0.3mmol, 33mg, CAS: 13881-91-9) in methanol / dichloromethane (V:V=2mL:2mL), stir at room temperature for 2h, then add sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg), and continue stirring. TLC is used to monitor the progress of the reaction. When 3a is consumed, PTLC separation (eluent is petroleum ether: ethyl acetate = 2:1) is performed to obtain compound II1 as a yellow solid of 16mg. The yield is 16%.

[0168] The reaction that occurs in this embodiment is shown in the following formula:

[0169]

[0170] The test results of the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula II1 is indeed obtained in this example.

[0171] NMR data of compound Ⅱ1, 1 H NMR (600 MHz, CDCl3-d 1)δ1.17-1.42(m,6H),1.69-1.78(m,6H),2.85-2.89(m,1H),4.74(s,2H),5.08(s,2 H),7.14-7.16(m,2H),7.37-7.42(m,2H),7.53(d,J=7.8Hz,1H),7.65-7.58(m,4H). 13 C NMR (150MHz, CDCl3-d 1 )δ25.07,25.78,33.52,38.96,38.97,64.50,68.28,106.03,118.18,123.87,123.91,123.95,123.99,124.54 ,124.94,126.26,126.90,127.10,127.44,127.99,128.54,129.99,132.97,133.28,135.28,145.92,155.65.

[0172] Embodiment 14

[0173] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminosulfonic acid compound, the specific structure of which is shown in Formula II2, and the specific process is as follows:

[0174] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0175] S2. Dissolve compound 3a (0.2mmol, 82mg) and taurine (0.3mmol, 38mg, CAS: 107-35-7) in methanol / dichloromethane (V:V=2mL:2mL), stir at room temperature for 2h, add sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg), and continue stirring. The purification method is the same as Example 13-S2 to obtain compound II2, a colorless oily solid of 54mg. The yield is 52%.

[0176] The reaction that occurs in this embodiment is shown in the following formula:

[0177]

[0178] The test results of the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula II2 is indeed obtained in this example.

[0179] NMR data of compound Ⅱ2, 1 H NMR (600 MHz, CDCl3-d 1)δ1.18-1.25(m,2H),1.32-1.42(m,4H),1.70-1.78(m,8H),2.85-2.89(m,1H),4.75(s,2H),5.08(s,2H),7.1 5-7.17(m,2H),7.38(dd,J=1.8,8.4Hz,1H),7.41(d,J=7.8Hz,1H),7.53(d,J=8.4Hz,1H),7.65-7.69(m,4H); 13 C NMR (150MHz, CDCl3-d 1 )δ25.07,25.78,33.52,38.96,64.52,68.29,106.03,118.19,122.62,123.91,123.95,124.44,124.54,12 4.94,126.27,126.91,127.10,127.44,128.00,128.54,129.99,132.97,133.28,135.29,145.92,155.66.

[0180] Embodiment 15

[0181] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminosulfonic acid compound, the specific structure of which is shown in Formula II3, and the specific process is as follows:

[0182] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0183] S2. Compound 3a (0.2mmol, 82mg) and homotaurine (0.3mmol, 42mg, CAS: 3687-18-1) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), stirred at room temperature for 2h, then sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added, and stirring was continued. The purification method was the same as that of Example 13-S2 to obtain compound II3 as a yellow solid (60mg), with a yield of 56%.

[0184] The reaction that occurs in this embodiment is shown in the following formula:

[0185]

[0186] The test results of the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula II3 is indeed obtained in this example.

[0187] NMR data of compound Ⅱ3, 1 H NMR (600 MHz, CDCl3-d1 )δ1.25-1.49(m,6H),1.58-1.67(m,4H),1.77-.86(m,6H),2.93-2.96(m,1H),4.82(s,2H),5.16(s,2H),7.2 2-7.24(m,2H),7.45(dd,J=1.2,8.4Hz,1H),7.48(d,J=7.8Hz,1H),7.60(d,J=84Hz,1H),7.72-7.76(m,4H); 13 CNMR (150MHz, CDCl3-d 1 )δ26.10,26.81,34.55,39.99,65.55,69.32.107.06,119.22,123.65,124.91,124.94,124.99,125.02,125.47,1 25.57,125.97,127.30,127.94,128.14,28.47,129.03,129.57,131.03,134.01,134.31,136.33,146.96,156.69.

[0188] Example 16

[0189] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminosulfonic acid compound, the specific structure of which is shown in Formula II4, and the specific process is as follows:

[0190] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0191] S2. Compound 3a (0.2mmol, 82mg) and 2-aminoethyl hydrogen sulfate (0.3mmol, 42mg, CAS: 926-39-6) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), stirred at room temperature for 2h, then sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added, and stirring was continued. The purification method was the same as that of Example 13-S2 to obtain compound II4 as a yellow solid of 10mg, with a yield of 10%.

[0192] The reaction that occurs in this embodiment is shown in the following formula:

[0193]

[0194] The test results of the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula II4 is indeed obtained in this example.

[0195] NMR data of compound Ⅱ4,1 H NMR (600 MHz, CDCl3-d 1 )δ1.18-1.23(m,4H),1.33-1.40(m,5H),1.70-1.78(m,5H),2.85-2.89(m,1H),4.75(s,2H),5.08(s,2H),7.1 5-7.17(m,2H),7.38(dd,J=1.2,8.4Hz,1H),7.41(d,J=8.4Hz,1H),7.53(d,J=7.8Hz,1H),7.65-7.69(m,4H); 13 C NMR (150MHz, CDCl3-d 1 )δ26.10,26.81,29.71,34.55,39.99,65.54,69.32,107.06,119.21,124.91,124.94,124.98,125.02,125.57 ,125.97,127.30,127.94,128.13,128.47,129.03,129.57,131.03,134.01,134.31,136.33,146.95,156.68.

[0196] Embodiment 17

[0197] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminophosphoric acid compound, the specific structure of which is shown in Formula II5, and the specific process is as follows:

[0198] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0199] S2. Compound 3a (0.2mmol, 82mg) and aminomethylphosphoric acid (0.3mmol, 33mg, CAS: 1066-51-9) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), stirred at room temperature for 2h, then sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added, and stirring was continued. The purification method was the same as that of Example 13-S2 to obtain compound II5 as a brown solid (32mg), with a yield of 32%.

[0200] The reaction that occurs in this embodiment is shown in the following formula:

[0201]

[0202] The test results of the aminophosphoric acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminophosphoric acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula II-5 is indeed obtained in this example.

[0203] NMR data of compound Ⅱ5, 1 H NMR (600 MHz, CDCl3-d 1 )δ1.26-1.50(m,6H),1.78-1.86(m,6H),2.94(t,J=9.6Hz,1H),4.81(s,2H),5.15(s, 2H),7.22-7.24(m,2H),7.45-7.50(m,2H),7.60(d,J=7.8Hz,1H),7.73-7.76(m,4H), 13 C-NMR (150MHz, CDCl3-d 1 )δ26.11,26.82,29.72,34.56,40.01,65.51,69.32,107.07,113.95,119.21,121.86,123.67,124.91,124.95,124.99,125.02,125.49,12 5.57,125.98,127.29,127.75,127.81,127.94,128.14,128.33,128. 48,129.03,129.58,131.04,134.01,134.33,136.34,146.96,159.69.

[0204] Embodiment 18

[0205] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminophosphoric acid compound, the specific structure of which is shown in Formula II6, and the specific process is as follows:

[0206] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0207] S2. Compound 3a (0.2mmol, 82mg) and aminoethylphosphoric acid (0.3mmol, 37mg, CAS: 2041-14-7) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), stirred at room temperature for 2h, then sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added, and stirring was continued. The purification method was the same as that of Example 13-S2 to obtain compound II6 as a colorless oily solid (57mg), with a yield of 55%.

[0208] The reaction that occurs in this embodiment is shown in the following formula:

[0209]

[0210] The test results of the aminophosphoric acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminophosphoric acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula II-6 is indeed obtained in this example.

[0211] NMR data of compound Ⅱ6, 1 H NMR (600 MHz, CDCl3-d 1 )δ1.28-1.47(m,6H),1.67-1.86(m,8H),2.93-2.96(m,1H),4.83(s,2H),5.16(s,2H),7.22-7.24(m ,2H),7.45(dd,J=1.8,8.4Hz,1H),7.48(d,J=8.4Hz,1H),7.60(d,J=7.8Hz,1H),7.72-7.76(m,4H); 13 C NMR (150MHz, CDCl3-d 1 )δ26.11,26.81,29.71,34.55,39.99,65.55,69.32,107.06,119.22,124.91,124.95,124.99,125.02,125.47,12 5.57,125.97,127.30,127.94,128.14,128.47,129.03,129.57,131.03,134.01,134.31,136.32,146.96,156.69.

[0212] Embodiment 19

[0213] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula III1, and the specific process is as follows:

[0214] S1. Dissolve 3-trifluoromethyl-4-cyclohexylbenzyl alcohol (5.0mmol, 1290mg), 4-hydroxybenzaldehyde (5.0mmol, 611mg, CAS: 123-08-0) and triphenylphosphine (PPh3, 6.0mmol, 1570mg) in anhydrous tetrahydrofuran (10mL), stir at room temperature, slowly add diisopropyl azodicarboxylate (DIAD, 6.0mmol, 1210mg) dropwise within 5min, and continue stirring. The reaction was allowed to react overnight at room temperature. After the reaction was completed, the reaction solvent was dried and separated by column chromatography (eluent was petroleum ether: ethyl acetate = 20:1) to obtain compound 3b, a white powdery solid 996mg, with a yield of 55%.

[0215] S2. Compound 3b (0.2mmol, 72mg) and glycine methyl ester hydrochloride (0.3mmol, 38mg) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), and N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added. After stirring at room temperature for 2h, sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added and continued to stir. TLC was used to monitor the progress of the reaction. When 3b was consumed, saturated sodium bicarbonate solution (20mL) was added, and then extracted with ethyl acetate, the organic phase was collected, concentrated under reduced pressure, and separated by PTLC (developing solvent was petroleum ether: ethyl acetate = 2:1) to obtain compound 4m as a colorless oily solid.

[0216] S3. The compound 4m obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound III1 as a white powder (82 mg). The yield was 98%.

[0217] The reaction that occurs in this embodiment is shown in the following formula:

[0218]

[0219] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula III1 is indeed obtained in this example.

[0220] NMR data of compound 3b, 1 H NMR (600 MHz, CDCl3-d 1 )δ1.28-1.30(m,1H),1.39-1.49(m,4H),1.77-1.86(m,5H),2.92-2.96(m,1H),5.13(s,2H),7.07(d,J= 9.0Hz,2H),7.48(d,J=8.4Hz,1H),7.55-7.56(m,1H),7.67(s,1H),7.85(d,J=9.0Hz,2H),9.90(s,1H); 13 C NMR (150MHz, CDCl3-d 1)δ26.08,26.78,34.53,40.01,69.53,115.08,123.55,124.84,124.88,124.92,124.96,125.37,128.07,128.26,128.60,130.35,130.94,132.05,133.43,147.32,163.46,190.75. High resolution mass spectrometry data: HRMS (ESI) calcd for C 21 H 21 F3O2(M+H) + :363.1567, found 363.1577.

[0221] NMR data of compound Ⅲ1, 1 H NMR (600 MHz, MeOD-d 4 )δ1.33-1.56(m,5H),1.76-1.87(m,5H),3.46(s,2H),4.13(s,2H),5.15(s,2H),7.06(d,J=8 .4Hz,2H),7.41(d,J=9.0Hz,2H),7.55(d,J=8.4Hz,1H),7.62(d,J=7.8Hz,1H),7.67(s,1H); 13 C NMR (150 MHz, MeOD-d 4 )δ25.67,26.57,34.17,40.06,50.01,68.64,115.14,123.71,123.81,124.03,124.07,124.11,124.15,125.62,127.23,127.42,128.32,130.93,131.21,135.11,146.35,159.43,169.31. High resolution mass spectrometry data: HRMS (ESI) calcd for C 23 H 26 F3NO3(M+H) + :422.1938,found422.1946.

[0222] Embodiment 20

[0223] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula III2, and the specific process is as follows:

[0224] S1. The synthesis of compound 3b was the same as Example 19-S1.

[0225] S2. Compound 3b (0.2mmol, 72mg) and dimethyl glutamate hydrochloride (0.3mmol, 64mg) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), and N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added. After stirring at room temperature for 2h, sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added and stirring was continued. The purification method was the same as that of Example 19-S2 to obtain compound 4n as a colorless oily solid.

[0226] S3. The compound 4n obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound III2 as a white powder (24 mg). The yield was 25%.

[0227] The reaction that occurs in this embodiment is shown in the following formula:

[0228]

[0229] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula III2 is indeed obtained in this example.

[0230] NMR data of compound Ⅲ2, 1 H NMR (600 MHz, MeOD-d 4 )δ1.35-1.56(m,5H),1.77-1.87(m,5H),2.05-2.09(m,1H),2.28-2.33(m,1H),2.37 -2.42(m,1H),2.48-2.54(m,1H),2.90(t,J=11.4Hz,1H),3.91(d,J=15.0Hz,1H),3. 96(dd,J=3.0,9.6Hz,1H),4.92(d,J=15.0Hz,1H),5.09(s,1H),6.96(d,J=8.4Hz,2H ),7.15(d,J=8.4Hz,2H),7.54(d,J=8.4Hz,1H),7.61(d,J=1.8Hz,1H),7.67(s,1H); 13 C NMR (150 MHz, MeOD-d 4)δ2.29,22.49,25.68,26.58,29.31,31.34,34.18,40.06,44.56,58.91,48.67,114.78,123.83,124.03,124.07,124.11,124.15,125.64,127.21,127.40,128.09,128.26,129.46,130.88,135.35,146.28,158.34,173.58,176.50. High resolution mass spectrometry data: HRMS (ESI) calcd for C 26 H 30 F3NO5(M-H2O+H) + :476.2043,found476.2052.

[0231] Embodiment 21

[0232] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula III3, and the specific process is as follows:

[0233] S1. The synthesis of compound 3b was the same as Example 19-S1.

[0234] S2. Compound 3b (0.2mmol, 72mg) and methionine methyl ester hydrochloride (0.3mmol, 60mg) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), and N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added. After stirring at room temperature for 2h, sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added and stirring was continued. The purification method was the same as that of Example 19-S2 to obtain compound 4o as a colorless oily solid.

[0235] S3. Compound 4o obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h and filtered to obtain compound III3 as a white powder (16 mg). The yield was 16%.

[0236] The reaction that occurs in this embodiment is shown in the following formula:

[0237]

[0238] The test results of the amino acid compounds substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compounds substituted with trifluoromethylcyclohexylbenzyl ether corresponding to formula III3 are indeed obtained in this example.

[0239] NMR data of compound Ⅲ3, 1 H NMR (600 MHz, MeOD-d 4 )δ1.24-1.47(m,5H),1.66-1.78(m,5H),1.97(d,J=3.0Hz,3H),1.98-2.02(m,2H),2.45-2.52(m,2H),2.80-2.84(m,1H),3.70(s,1H),3. 95-4.08(m,2H),5.05(d,J=3.0Hz,2H),6.94-6.97(m,2H),7.26-7.33(m,2H),7.45(d,J=7.8Hz,1H),7.52(d,J=8.4Hz,1H),7.57(s,1H); 13 C NMR (150 MHz, MeOD-d 4 )δ13.70,25.67,26.56,29.21,29.84,34.18,40.05,49.84,50.15,58.25,60.74,68.59,115.13,123.65,124.06,124.10,128.31,130.90,131.37,146.35,159.43,171.26. High resolution mass spectrometry data: HRMS (ESI) calcd for C 26 H 32 F3NO3S(M+H) + :496.2128,found496.2128.

[0240] Embodiment 22

[0241] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound, the specific structure of which is shown in Formula III4, and the specific process is as follows:

[0242] S1. The synthesis of compound 3b was the same as Example 19-S1.

[0243] S2. Compound 3b (0.2mmol, 72mg) and threonine methyl ester hydrochloride (0.3mmol, 51mg) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), N,N-diisopropylethylamine (DIPEA, 0.3mmol, 39mg) was added, and sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added after stirring at room temperature for 2h, and stirring was continued. The purification method was the same as that of Example 19-S2 to obtain compound 4p as a colorless oily solid.

[0244] S3. The compound 4p obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added, and stirred at room temperature for 3 h. After the reaction, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was continued to be added until a precipitate was formed. The mixture was allowed to stand for 10 h, and compound III4 was obtained by filtration as a white powder (54 mg) with a yield of 58%.

[0245] The reaction that occurs in this embodiment is shown in the following formula:

[0246]

[0247] The test results of the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula III4 is indeed obtained in this example.

[0248] NMR data of compound Ⅲ4, 1 H NMR (600 MHz, DMSO-d 6 )δ1.15(d,J=6.0Hz,3H),1.30-1.56(m,5H),1.71-1.84(m,5H),2.83(t,J =11.4Hz,1H),3.09(t,J=6.6Hz,1H),3.91-3.95(m1H),4.01(d,J=13.2Hz ,1H),4.10(d,J=13.2Hz,1H),5.16(s,2H),7.04(d,J=9.0Hz,2H),7.43(d ,J=9.0Hz,2H),7.61(d,J=7.8Hz,1H),7.67(d,J=8.4Hz,1H),7.71(s,1H); 13 C NMR (150 MHz, DMSO-d 6 )δ20.89,25.91,26.90,34.40,49.58,65.90,66.29,68.79,115.21,124.19,124.82,124.86,124.90,126.00,126.90,127.10,129.02,132.08,135.65,146.48,159.17,169.32. High resolution mass spectrometry data: HRMS (ESI) calcd for C 25 H 30 F3NO4(M+H) + :466.2200,found466.2205.

[0249] Embodiment 23

[0250] This example prepares a trifluoromethylcyclohexylbenzyl ether substituted aminosulfonic acid compound, the specific structure of which is shown in Formula IV1, and the specific process is as follows:

[0251] S1. The synthesis of compound 3b was the same as Example 19-S1.

[0252] S2. Compound 3b (0.2mmol, 72mg) and taurine (0.3mmol, 38mg) were dissolved in methanol / dichloromethane (V:V=2mL:2mL), stirred at room temperature for 2h, then sodium cyanoborohydride (0.3mmol, 19mg) and acetic acid (0.8mmol, 48mg) were added, and stirring was continued. The purification method was the same as Example 13-S2 to obtain compound IV1, 52mg of yellow oily solid, with a yield of 56%.

[0253] The reaction that occurs in this embodiment is shown in the following formula:

[0254]

[0255] The test results of the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether obtained in this example are as follows, indicating that the aminosulfonic acid compound substituted with trifluoromethylcyclohexylbenzyl ether corresponding to Formula IV1 is indeed obtained in this example.

[0256] NMR data of compound Ⅳ1, 1 H NMR (600 MHz, CDCl3-d 1 )δ1.27-1.48(m,7H),1.77-1.86(m,6H),1.97(s,1H),2.94(d,J=9.6Hz,1H),4.62(s,2H),5.04(s ,2H),6.96(d,J=9.0Hz,2H),7.29(d,J=8.4Hz,2H),7.46(d,J=7.8Hz,1H),7.55(d,J=8.4Hz,1H); 13 C NMR (150MHz, CDCl3-d 1 )δ26.10,26.81,34.55,39.98,39.99,64.99,69.33,114.87,123.65,124.76,124.80,124.83,124.88,125.47,127.90,128.09,128.44,128.72,130.90,133.65,134.44,146.88,158.20.Pharmacological experiments

[0257] Example 1 HTRF-IP1 method to detect the in vitro agonist activity of compounds on S1P1 and S1P3

[0258] cell:

[0259] Transfected Chinese hamster ovary-K1 (CHO-K1) Gαqi5 cells hS1P1-CHO with stable high expression of S1P1 were constructed by Multispan (Hayward, California, USA). The cells were cultured in DMED / F12 medium containing 10 μg / mL puromycin, 250 μg / mL hygromycin and 10% fetal bovine serum, and passaged 1:3.

[0260] drug:

[0261] 1×Stimb Buffer (containing 1% BSA without free fatty acids) was used to prepare different concentrations of compounds I1-12, II1-6, III1-4, IV1 and the positive drug FTY720-P.

[0262] instrument:

[0263] EnVision Multi-function Microplate Reader (PerkinElmer)

[0264] method:

[0265] The homogeneous time-resolved fluorescence-IP1 (HTRF-IP1) method was used to detect the concentration of IP1 produced after S1P1 or S1P3 stimulation and calculate the EC 50 The value of this reaction compound is the agonist activity of the receptor. IP1 was detected using the IP1 kit (Cisbio). First, hS1P1-CHO and hS1P3-CHO cell buffers were prepared using 1×Stimb Buffer, and 7 μL of 1×10 7 100 μL of cells / mL. Then, 7 μL of compounds of different concentrations were added to the plate and incubated at 37°C and 5% CO2 for 2 hours. Then, IP1-d2 and Ab-Cryp were prepared with 1×Lysis Buffer and 3 μL was added to each well. After incubation at room temperature for 1 hour, the absorbance at 665 and 615 nm was detected using EnVision. The absorbance 665nm / 615nm (A 665 / A 615 ). The ratio (A 665 / A 615 ) and IP1 concentration standard curve. Data statistics:

[0266] The IP1 concentration of the test compound was calculated using the standard curve. 50 The values ​​were calculated using Graphpad software 8.0 (Graphpad, La Jolla, CA, USA).

[0267] Experimental results:

[0268] The results are shown in Table 1.

[0269] Table 1. Agonist activity of compounds on S1P1 and S1P3

[0270]

[0271]

[0272] Experimental conclusion:

[0273] 1) Positive drug FTY720-P has strong agonist activity on S1P1 and S1P3, EC 50 55.63nM and 43.27nM respectively.

[0274] 2) Compounds I6, I11, I12, II3, II6, and III4 have strong agonist activity on S1P1 but no agonist activity on S1P3.

[0275] Example 2 Effects of the Detection Compound on Rat Peripheral Lymphocytes

[0276] animal:

[0277] Sprague-Dawley rats, 140-160 g, male

[0278] Drugs: Use CMC-Na to prepare FTY720, I6, I11, I12, II3, and II6 of different concentrations.

[0279] Instrument: Five-differentiation blood cell analyzer (MEK-7222K, Nihon Kohden)

[0280] Experimental groups:

[0281] Normal control group;

[0282] Positive control group: FTY720 (3 mg / kg) group;

[0283] Test compound groups: I6 (3 mg / kg), I11 (3 mg / kg), I12 (0.3 mg / kg), I12 (1 mg / kg), I12 (3 mg / kg), I12 (10 mg / kg), II3 (3 mg / kg), and II6 (3 mg / kg) groups.

[0284] method:

[0285] First, the rats were randomly divided into groups, with 5 rats in each group. 3 mg / kg of the positive drug FTY720, 3 mg / kg of I6, 3 mg / kg of I11, 0.3 mg / kg of I12, 1 mg / kg of I12, 3 mg / kg of I12, 10 mg / kg of I12, 3 mg / kg of II3, 3 mg / kg of II6 or a control solute (0.5% CMC-Na) were administered by gavage. Blood (20 μL) was collected from the tail vein at 0, 2, 4, 8, 12, 24 and 48 hours after administration, and peripheral blood lymphocytes were counted using a hematology analyzer (10 9 / L).

[0286] Statistics:

[0287] The maximum percentage of lymphocyte reduction in each group was calculated, and the data of each group were compared between groups using T.TEST. P<0.05 was considered to be significantly different.

[0288] Experimental results:

[0289] The results are shown in Table 2 and Table 3. Figure 1 and Figure 2 .

[0290] Table 2. Maximum reduction percentage of rat lymphocyte number by compounds

[0291]

[0292]

[0293]

[0294] Compared with the control group, *P<0.05, ***P<0.001.

[0295] Table 3. Maximum reduction percentage of rat lymphocyte number by different concentrations of I12

[0296] Compared with the control group, *P 0.05, ***P < 0.001.

[0297] Experimental conclusion:

[0298] 1) The positive drug FTY720 significantly reduced the number of peripheral blood lymphocytes in rats after administration, P<0.001, with statistical significance.

[0299] 2) Among the tested compounds, I12 had the strongest effect on reducing the expression of rat peripheral blood lymphocytes and had a good dose-dependency, P<0.05, which was statistically significant.

[0300] Example 3 Detection of the effect of compound I12 on mouse pulmonary fibrosis model

[0301] animal:

[0302] C57BL / 6J mice, 6-8 weeks, 20-22 g, male.

[0303] Drugs: Nintedanib (MedChemExpress, HY-50904 / CS-0104); Bleomycin hydrochloride for injection (Nippon Kayaku Co., Ltd., 600700); I12 of different concentrations was prepared using CMC-Na.

[0304] Instrument: Mouse pulmonary function tester (flexiVent TM system, Scireq), electronic balance.

[0305] Experimental groups:

[0306] Normal control group;

[0307] Model group;

[0308] Positive control group: nintedanib (50 mg / kg) group;

[0309] Test compound groups: I12 (0.3 mg / kg), I12 (1 mg / kg), and I12 (3 mg / kg) groups.

[0310] method:

[0311] First, the mice were randomly divided into groups, with 14 mice in the control group and 78 mice in the modeling group. A mouse pulmonary fibrosis model was established by a single tracheal instillation of bleomycin 2.5 mg / kg. After the 10th day, the modeled mice were randomly divided into groups, with 16 mice in the model group and 13 mice in the drug administration group. The mice in the drug administration group were gavaged with 50 mg / kg of the positive drug nintedanib, 0.3 mg / kg of I12, 1 mg / kg of I12, and 3 mg / kg of I12 every day, and the model group was given a control solute (0.5% CMC-Na). On the 31st day, the mice were anesthetized and the lung function was detected using a mouse spirometer (n=5).

[0312] Statistics:

[0313] Four indicators of lung function were measured: dynamic compliance Crs, dynamic resistance Rrs, dynamic elasticity Ers and deep inspiratory capacity IC. The data of each group were compared among the groups using T.TEST. P<0.05 was considered to be significantly different.

[0314] Experimental results:

[0315] Results Figure 3-Figure 6 .

[0316] Experimental conclusion:

[0317] 1) Single intratracheal instillation of bleomycin induced deterioration of lung function in mice. Ers index of mice increased significantly. Crs and IC decreased significantly, while Rrs did not change significantly.

[0318] 2) The positive drug nintedanib significantly improved the lung function of fibrotic mice. Crs and IC increased significantly, and Ers index decreased significantly.

[0319] 3) All concentrations of compound I12 significantly improved the lung function of fibrotic mice, with Crs and IC increased significantly and Ers decreased significantly in a dose-dependent manner.

[0320] Example 4 Detection of the effect of compound I12 on acute colitis in mice

[0321] animal:

[0322] C57BL / 6J mice, 6-8 weeks, 20-22 g, male.

[0323] Drugs: 2% dextran sulfate sodium DSS (MD, item number 160110, batch number S8634); CMC-Na was used to prepare different concentrations of I12.

[0324] Instruments: ruler, electronic balance.

[0325] Experimental groups:

[0326] Normal control group;

[0327] Model group;

[0328] Test compound groups: I12 (0.3 mg / kg), I12 (1 mg / kg), and I12 (3 mg / kg) groups.

[0329] method:

[0330] First, the mice were randomly divided into groups, with 11 mice in the control group, 12 mice in the model group, I12 (0.3 mg / kg), I12 (1 mg / kg), and I12 (3 mg / kg) groups. The mouse acute colitis model was established by giving the mice free drinking of 2% DSS aqueous solution for 7 consecutive days. On the day of modeling, 0.3 mg / kg of I12, 1 mg / kg of I12, and 3 mg / kg of I12 were gavaged, and the model group was given a control solute (0.5% CMC-Na). After 7 days, the disease activity index DAI of the mice was evaluated, the mice were over-anesthetized, the mouse colon was taken, the length was measured and photographed.

[0331] Statistics:

[0332] The three indicators of DAI, weight loss percentage, stool shape and degree of blood in stool, were measured. The scoring criteria are shown in Table 4:

[0333] Table 4. Scoring criteria for the disease activity index of colitis in mice

[0334]

[0335] Note: ①Normal stool: formed stool; ②Loose stool: pasty, semi-formed stool that does not adhere to the anus; ③Loose stool: loose stool

[0336] T.TEST was used to compare the data among groups, and P < 0.05 was considered to be a significant difference.

[0337] Experimental results:

[0338] Results Figure 7-Figure 12 .

[0339] Experimental conclusion:

[0340] 1) 2% DSS model successfully induced acute colitis in mice. The length of the colon of mice was significantly shortened, the DAI scores were significantly increased, the body weight decreased significantly, and obvious bloody and loose stools appeared.

[0341] 2) All concentrations of compound I12 significantly improved the colon length of colitis mice, and the DAI scores were significantly reduced compared with the model group, the weight loss was reduced, and the bloody and loose stools were significantly reduced, with a certain dose-dependency.

Claims

1. A nitrogen-containing organic acid derivative substituted with trifluoromethylcyclohexylbenzyl ether, characterized in that: Including compounds represented by formula (I) and pharmaceutically acceptable salts thereof: in, R1 is selected from one of phenyl and naphthyl; R2 is selected from One of; R3 is selected from one of H and methyl.

2. The trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative according to claim 1, characterized in that: The phenyl and naphthyl groups are: One of them.

3. The trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative according to any one of claims 1 or 2, characterized in that: The compound is selected from the following:

4. A method for synthesizing the trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative according to any one of claims 1 to 3, characterized in that: The steps include: (1) Using 3-trifluoromethyl, 4-cyclohexylbenzyl alcohol represented by formula (Ia) as a substrate and 6-OH, 2-naphthaldehyde or 4-OH benzaldehyde for Mitsunobu reaction to obtain; (2) the aldehyde group of formula (Ib) reacts with the amino group of NH2R to obtain a nitrogen-containing sulfonic acid / phosphoric acid compound substituted with trifluoromethylcyclohexylbenzyl ether as shown in formula (I); (3) the aldehyde group of formula (Ib) reacts with the amino group of NH2R to obtain an amino acid ester compound substituted with trifluoromethylcyclohexylbenzyl ether as shown in formula (Ic); (4) The compound of formula (Ic) is hydrolyzed in an aqueous solution of lithium hydroxide to obtain a trifluoromethylcyclohexylbenzyl ether substituted amino acid compound represented by formula (I); 5. The synthesis method according to claim 4, characterized in that The reaction of step (1) is carried out in tetrahydrofuran or toluene at room temperature, and diisopropyl azodicarboxylate and triphenylphosphine are added as catalysts. The molar ratio of benzyl alcohol, 6-hydroxy, 2-naphthaldehyde or 4-hydroxybenzaldehyde, diisopropyl azodicarboxylate and triphenylphosphine is 1.0:1.0:1.2:1.

2.

6. The synthesis method according to claim 4, characterized in that The reaction in step (2) is carried out in methanol / dichloromethane (V:V=1:1) at room temperature, and after stirring for 2 hours, a reducing agent, sodium cyanoborohydride and acetic acid are added, wherein R is selected from alkyl-substituted sulfonic acid and phosphoric acid, and the molar ratio of the formula (Ib), NH2R, sodium cyanoborohydride and acetic acid is 1.0:1.5:4.0:1.

0.

7. The synthesis method according to claim 4, characterized in that The reaction of step (3) is carried out in methanol / dichloromethane (V:V=1:1) at room temperature, and N,N-diisopropylethylamine is added at the same time. After stirring for 2 hours, a reducing agent, sodium cyanoborohydride and acetic acid are added. R is selected from amino acid ester hydrochlorides of different substitutions, and the molar ratio of the formula (Ib), NH2R, N,N-diisopropylethylamine, sodium cyanoborohydride and acetic acid is 1.0:1.5:1.5:4.0:1.

0.

8. The synthesis method according to claim 4, characterized in that The reaction of step (4) is carried out in methanol, 0.5 mol / L lithium hydroxide aqueous solution is added, the reaction temperature is room temperature, and the molar ratio of the formula (Ic) to lithium hydroxide is 1.0:8.

0.

9. Use of the trifluoromethylcyclohexylbenzyl ether substituted nitrogen-containing organic acid derivative according to any one of claims 1 to 3 in the preparation of drugs for preventing or treating inflammation-related diseases.

10. The use according to claim 9, characterized in that The inflammation-related diseases include pulmonary fibrosis, ulcerative colitis, multiple sclerosis, etc.

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