A trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative, its preparation and use

Abstract

The application discloses a trifluoromethyl cyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative, 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 is obtained through two-step reaction, that is, first, a light extension reaction is carried out on a benzyl alcohol compound and hydroxynaphthaldehyde (or hydroxybenzaldehyde), then a Schiff base is generated after the amino acid ester hydrochloride (or amino sulfonic acid, or amino phosphoric acid) is reacted with the benzyl alcohol compound, and sodium cyanoborohydride is used for reduction (or simple hydrolysis is carried out after reduction), so the trifluoromethyl cyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative is obtained. The preparation method 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 adjust S1P1 receptors without exciting S1P3 receptors, and is expected to be developed into a new preparation for treating inflammatory and immune related diseases such as pulmonary fibrosis, ulcerative colitis and multiple sclerosis.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of organic compound synthesis and medicine, and particularly relates to a trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative, a synthesis method thereof and pharmaceutical use. BACKGROUND

[0002] Sphingosine-1-phosphate (S1P) is an endogenous ligand for a family of five G protein-coupled receptors (S1P 1-5 ) on the surface of cell membranes, which can regulate a variety of biological processes, including cell differentiation, vascular stability, inflammation, endothelial integrity and angiogenesis. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid substance 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 stimulating S1P3 can be a new way to treat idiopathic pulmonary fibrosis (IPF), ulcerative colitis, multiple sclerosis and other inflammation-related diseases.

[0003] Therefore, exploring the structural characteristics of S1P1, mimicking the binding mode of the compound and the receptor, and targeting design and synthesis of new selective S1P1 modulators are expected to find effective drugs for treating pulmonary fibrosis, ulcerative colitis, multiple sclerosis and other inflammation-related diseases. SUMMARY

[0004] In view of the above technical problems, the present application designs a series of trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivatives. 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, an aminosulfonic acid or an aminophosphonic acid as a hydrophilic polar head. The compounds can selectively regulate S1P1 receptors and have high application value in the medical field.

[0005] The present application also provides a synthesis method of the trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative.

[0006] The present application also provides a pharmaceutical use of the trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative.

[0007] According to an aspect of the present application, a trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivative is provided, which includes a compound having a structure shown in formula (I) and a pharmaceutically acceptable salt thereof:

[0008]

[0009] In the formula,

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

[0011] R2 is selected from

[0012] one of the following:

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

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

[0015] one of the following:

[0016] In some embodiments of the present application, when R1 is selected from a benzene ring in the formula (I), R3 is selected from hydrogen, and R2 is selected from one of the following:

[0017] In some embodiments of the present application, at least one of the following compounds of the trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivatives 11-12, 11-6, 111-4, IV1 is shown in the structure:

[0018]

[0019]

[0020]

[0021] According to another aspect of the present application, a synthesis method of the trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid derivatives is provided, which comprises first performing a photo extension reaction of a benzyl alcohol compound of formula (Ia) with a hydroxynaphthaldehyde (or hydroxybenzaldehyde) to generate formula (Ib), then generating a Schiff base with an amino acid (or amino sulfonic acid, or amino phosphoric acid), and then reducing with sodium cyanoborohydride to obtain the trifluoromethylcyclohexyl benzyl ether substituted nitrogen-containing organic acid compound of formula (I).

[0022]

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

[0024]

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

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

[0027] In some embodiments of the present application, 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 application, the reaction temperature for steps (2) and (3) is room temperature (5-30℃).

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

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

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

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

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

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

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

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

[0037] Beneficial technical effects

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

[0039] 2. The application proves that the selectivity of compounds I6, I11, I12, II3, II6 and III4 is higher, and the compounds have stronger agonistic activity on S1P1 and no agonistic activity on S1P3 through the test research on the S1P1 agonistic activity of the nitrogen-containing organic acid derivative substituted by trifluoromethylcyclohexyl benzyl ether;

[0040] 3. The application proves that I12 has obvious anti-fibrosis effect through the pharmacodynamic test research of the nitrogen-containing organic acid derivative substituted by trifluoromethylcyclohexyl benzyl ether on mouse pulmonary fibrosis;

[0041] 4. The application proves that I12 has obvious anti-colitis effect through the pharmacodynamic test research of the nitrogen-containing organic acid derivative substituted by trifluoromethylcyclohexyl benzyl ether on mouse colitis; BRIEF DESCRIPTION OF DRAWINGS

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

[0043] Figure 2 Effect of I12 at different concentrations on rat peripheral blood lymphocytes (12 hours), compared with the control group, *P<0.05, ***P<0.001;

[0044] Figure 3 Effect of I12 at different concentrations on mouse lung dynamic compliance, compared with the control group, ####P<0.0001; compared with the model group, *P<0.05, ***P<0.001;

[0045] Figure 4 Effect of I12 at different concentrations on mouse lung dynamic resistance, compared with the control group, #P<0.05; compared with the model group, *P<0.05;

[0046] Figure 5 Effect of I12 at different concentrations on mouse dynamic elasticity, compared with the control group, ##P<0.01; compared with the model group, *P<0.05, **P<0.01;

[0047] Figure 6 Effect of I12 at different concentrations on mouse deep inhalation volume, compared with the control group, ####P<0.0001; compared with the model group, *P<0.05, **P<0.01;

[0048] Figure 7 Representative pictures of the effect of I12 at different concentrations on mouse colon length;

[0049] Figure 8.I12The influence of each concentration on the colon length of mice was compared with the control group, and the difference was statistically significant (####P<0.0001); compared with the model group, *P<0.05;

[0050] Figure 9 .I12The influence of each concentration on the colon length of mice was compared with the control group, and the difference was statistically significant (####P<0.0001); compared with the model group, *P<0.05;

[0051] Figure 10 .I12The influence of each concentration on the colon length of mice was compared with the control group, and the difference was statistically significant (####P<0.0001); compared with the model group, *P<0.05;

[0052] Figure 11 .I12The influence of each concentration on the colon length of mice was compared with the control group, and the difference was statistically significant (####P<0.0001); compared with the model group, *P<0.05;

[0053] Figure 12 .I12The influence of each concentration on the colon length of mice was compared with the control group, and the difference was statistically significant (####P<0.0001); compared with the model group, *P<0.05; DETAILED DESCRIPTION

[0054] The present application will be described in detail below with reference to examples, so that the purpose, features and effects of the present application can be fully understood. Obviously, the described examples are only some of the examples of the present application, but not all the examples. Based on the examples of the present application, other examples obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0055] Example 1

[0056] In this example, a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound is prepared, and the specific structure is shown as formula I1, and the specific process is as follows:

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

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

[0059] S3. Compound 4a 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, CAS: 1310-65-2) was added, and the stirring was continued at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid (CAS: 7647-01-0) was added to adjust the pH to 1-2, and water was further added until a precipitate was precipitated, and the mixture was allowed to stand for 10 h, and then filtered to obtain compound I1 as a white powder 60 mg with a yield of 64%.

[0060] The reaction of this example occurs as shown in the following formula:

[0061]

[0062] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula II is indeed obtained in this example.

[0063] NMR data of compound 3a, 1 H NMR (600 MHz, MeOD-d 1 ) δ 1.25-1.33 (m, 1H), 1.39-1.49 (m, 4H), 1.77 (d, J = 12.6 Hz, 1H), 1.82-1.86 (m, 4H), 2.93-2.97 (m, 1H), 7.26 (d, J = 2.4 Hz, 1H), 7.30 (dd, J = 2.4, 9.0 Hz, 1H), 7.50 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.72 (s, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.91-7.93 (m, 2H), 8.26 (s, 1H), 10.10 (s, 1H); 13 C NMR (150 MHz, MeOD-d 1 ) δ 26.08, 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, found 413.1728.

[0064] NMR data of compound II, 1 H NMR (600 MHz, MeOD-d 4) δ 1.33-1.45 (m, 3H), 1.51-1.57 (m, 2H), 1.77 (d, J = 12.0 Hz, 3H), 1.85-1.87 (m, 2H), 2.91 (t, J = 11.4 Hz, 2H), 3.94 (s, 2H), 4.39 (s, 2H), 5.24 (s, 2H), 7.29 (dd, J = 2.4, 9.0 Hz, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.53 (dd, J = 0.6, 8.4 Hz, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.46 (s, 1H), 7.85-7.88 (m, 2H), 7.94 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 27.12, 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 spectral data: HRMS (ESI) calcd for C 27 H 28 F3NO3 (M+H) + : 472.2094, found 472.2097.

[0065] Example 2

[0066] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure is shown as formula 1 2, the specific process is:

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

[0068] S2. Compound 3a (0.2 mmol, 82 mg) was dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL) with aspartic acid methyl ester hydrochloride (0.3 mmol, 60 mg, CAS: 69630-50-8), N, N-diisopropyl ethylamine (DIPEA, 0.3 mmol, 39 mg) was added, after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and stirring was continued. The reaction progress was detected by TLC, and the purification method was the same as that in Example 1-S2 to obtain compound 4b, 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, and stirring was continued at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated. After standing for 10 h, filtration was performed to obtain compound I2, a white powder 27 mg, with a yield of 26%.

[0070] The reaction of this example is shown in the following formula:

[0071]

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

[0073] NMR data of compound I2, 1 H NMR (600 MHz, DMSO-d 6 ) δ 1.30-1.35 (m, 3H), 1.52-1.56 (m, 2H), 1.69 (d, J = 11.4 Hz, 3H), 1.80 (d, J = 12.0 Hz, 2H), 2.60-2.64 (m, 1H), 2.72 (dd, J = 9.0, 16.2 Hz, 1H), 2.80 (t, J = 11.4 Hz, 1H), 3.68-3.70 (m, 1H), 4.10 (d, J = 13.2 Hz, 1H), 4.19 (d, J = 13.2 Hz, 1H), 5.27 (s, 2H), 7.26-7.28 (m, 1H), 7.45 (d, J = 1.8 Hz, 1H), 7.52 (d, J = 8.4 Hz, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.79 (s, 1H), 7.78-7.87 (m, 3H); 13 C NMR (150 MHz, DMSO-d 6) δ 25.85, 26.88, 34.31, 36.73, 39.56, 39.70, 39.84, 39.97, 40.11, 40.25, 40.39, 50.14, 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 spectral data: HRMS (ESI) calcd for C 29 H 30 F3NO5(M+H) + :530.2149, found 530.2158.

[0074] Example 3

[0075] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure as shown in formula 13, the specific process is:

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

[0077] S2. Compound 3a (0.2 mmol, 82 mg) and leucine methyl ester hydrochloride (0.3 mmol, 60 mg, CAS: 7517-19-3) were dissolved in methanol / dichloromethane (V:V=2 mL:2 mL), N,N-diisopropylethylamine (DIPEA, 0.3 mmol, 39 mg) was added, and after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. TLC was used to detect the progress of the reaction, and the purification method was the same as example 1-S2 to obtain compound 4c, 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, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated. After standing for 10 h, filtration gave compound 13, white powder 89 mg, yield 85%.

[0079] The reaction occurred in this example is shown in the following formula:

[0080]

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

[0082] NMR data of compound I3, 1 H NMR (600 MHz, MeOD-d 4 ) δ 0.92 (d, J = 6.0 Hz, 1H), 0.97 (d, J = 6.0 Hz, 1H), 1.34-1.57 (m, 5H), 1.66-1.71 (m, 1H), 1.77-1.87 (m, 7H), 2.91 (t, J = 11.6 Hz, 1H), 3.75-3.78 (m, 1H), 4.28-4.35 (m, 2H), 5.22 (s, 3H), 7.27 (d, J = 9.0 Hz, 1H), 7.37 (s, 1H), 7.52 (d, J = 8.4 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.73 (s, 1H), 7.83-7.86 (m, 2H), 7.91 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 20.77, 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] A trifluoromethylcyclohexyl benzyl ether substituted amino acid compound is prepared in this example, and the specific structure is shown as formula I4, and the specific process is as follows:

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

[0086] S2. Compound 3a (0.2 mmol, 82 mg) was dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL) with proline methyl ester hydrochloride (0.3 mmol, 50 mg, CAS: 2133-40-6), N, N-diisopropyl ethylamine (DIPEA, 0.3 mmol, 39 mg) was added, after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and stirring was continued. The reaction progress was detected by TLC, and compound 4d was obtained by purification method same as example 1-S2, 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, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was added to precipitate, and the mixture was stirred for 10 h. The precipitate was filtered to obtain compound I4, yellow solid 57 mg, yield 56%.

[0088] The reaction of this example is shown in the following formula:

[0089]

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

[0091] NMR data of compound I4, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.32-1.55 (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 = 12.6 Hz, 1H), 4.58 (d, J = 12.6 Hz, 1H), 5.22 (s, 2H), 7.27 (d, J = 8.4 Hz, 1H), 7.36 (s, 1H), 7.57-7.58 (m, 2H), 7.67 (d, J = 7.2 Hz, 1H), 7.75 (s, 1H), 7.83 (d, J = 7.8 Hz, 2H), 7.96 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4) δ 12.00, 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 spectral data: HRMS (ESI) calcd for C 29 H 32 F3NO4(M+H) + :512.2407, found512.5037.

[0092] Example 5

[0093] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure as shown in formula 15, the specific process is:

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

[0095] S2. Compound 3a (0.2 mmol, 82 mg) and sarcosine methyl ester hydrochloride (0.3 mmol, 42 mg, CAS:13515-93-0) were dissolved in methanol / dichloromethane (V:V=2 mL:2 mL), N,N-diisopropylethylamine (DIPEA, 0.3 mmol, 39 mg) was added, and after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. TLC was used to detect the progress of the reaction, and the purification method was the same as example 1-S2 to obtain compound 4e, 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, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated. After standing for 10 h, filtration was carried out to obtain compound 15, a white oily substance 81 mg, with a yield of 84%.

[0097] The reaction occurred in this example is shown in the following formula:

[0098]

[0099] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula 15 is indeed obtained in this example.

[0100] Compound 15 NMR data, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.34-1.58 (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, 9.0 Hz, 1H), 7.41 (d, J = 2.4 Hz, 1H), 7.57-7.59 (m, 2H), 7.69 (d, J = 8.4 Hz, 1H), 7.75 (s, 1H), 7.88 (t, J = 9.0 Hz, 1H), 8.00 (s, 3H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 7.12, 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 spectral data: HRMS (ESI) calcd for C 28 H 30 F3NO3 (M+H) + : 486.2251, found 486.2259.

[0101] Example 6

[0102] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure is shown as formula 16, the specific process is:

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

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

[0105] S3. Compound 4f obtained in S2 was dissolved in methanol (4 mL), and lithium hydroxide aqueous solution (0.5 mol / L, 3.2 mL) was added, and stirred at room temperature for 3 h, after the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was added to precipitate, and the mixture was stirred for 10 h, and then filtered to obtain compound I6, yellow solid 54 mg, yield 50%.

[0106] The reaction of this example is shown in the following formula:

[0107]

[0108] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which shows that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula I6 is indeed obtained in this example.

[0109] NMR data of compound I6, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.34-1.57 (m, 5H), 1.77-1.88 (m, 5H), 2.06-2.11 (m, 1H), 2.30-2.35 (m, 1H), 2.42-2.47 (m, 1H), 2.52-2.58 (m, 1H), 2.91 (t, J = 11.4 Hz, 1H), 4.0 (dd, J = 3.0, 9.6 Hz, 1H), 4.12 (d, J = 15.0 Hz, 1H), 5.10 (d, J = 15.0 Hz, 1H), 5.21 (s, 2H), 7.21 (dd, J = 2.4, 9.0 Hz, 1H), 7.29-7.32 (m, 2H), 7.56 (d, J = 8.4 Hz, 1H), 7.63 (s, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.74-7.76 (m, 3H); 13 C NMR (150 MHz, MeOD-d 4) δ 24.01, 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 spectral data: HRMS (ESI) calcd for C 30 H 32 F3NO5(M-H2O+H) + :526.2200, found 526.2208.

[0110] Example 7

[0111] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure as shown in formula 17, the specific process is:

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

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

[0114] S3. Compound 4g obtained in S2 was dissolved in methanol (4 mL), and aqueous lithium hydroxide solution (0.5 mol / L, 3.2 mL) was added, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated, and then it was placed for 10 h, and then filtered to obtain compound 17, white powder 100 mg, yield 90%.

[0115] The reaction occurred in this example is shown in the following formula:

[0116]

[0117] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula I7 is indeed obtained in this example.

[0118] NMR data of compound I7, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.77-1.85 (m, 5H), 2.92-2.95 (m, 1H), 3.28-3.32 (m, 1H), 3.55 (d, J = 5.4, 1H), 3.84-3.86 (m, 1H), 4.13 (d, J = 13.2 Hz, 1H), 4.23 (m, 1H), 4.95 (s, 2H), 6.99 (d, J = 2.4 Hz), 7.12-7.14 (m, 3H), 7.21-7.23 (m, 3H), 7.47 (d, J = 7.8 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.66-7.68 (m, 3H), 7.70 (d, J = 9.0 Hz, 1H), 7.78 (S, 1H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 26.08, 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, 127.54, 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. High resolution mass spectrometry data: HRMS (ESI) calcd for C 34 H 34 F3NO3 (M+H) + : 562.2564, found 562.2576.

[0119] Example 8

[0120] A trifluoromethylcyclohexyl benzyl ether substituted amino acid compound is prepared in this example, and the specific structure is shown as formula I8, and the specific process is as follows:

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

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

[0123] S3. Compound 4h 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 stirring was continued at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated, and the mixture was allowed to stand for 10 h. Filtration gave compound I8, a white powder 39 mg, in a yield of 38%.

[0124] The reaction in this example occurred as shown in the following formula:

[0125]

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

[0127] NMR data of compound I8, 1 H NMR (600 MHz, MeOD-d 4 ) δ 0.94-0.98 (m, 6H), 1.37-1.57 (m, 5H), 1.77-1.88 (m, 5H), 2.07-2.08 (m, 1H), 2.91 (t, J = 11.4 Hz, 1H), 3.72 (s, 2H), 4.05 (d, J = 3.6 Hz, 1H), 4.39 (s, 2H), 5.24 (s, 2H), 7.29-7.31 (m, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.53-7.59 (m, 2H), 7.68 (d, J = 7.8 Hz, 1H), 7.74 (s, 1H), 7.85-7.87 (m, 2H), 7.94 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4) δ 12.05, 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 spectral data: HRMS (ESI) calcd for C 30 H 34 F3NO3(M+H) + :514.2564, found 514.2568.

[0128] Example 9

[0129] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure as shown in formula 19, the specific process is:

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

[0131] S2. Compound 3a (0.2 mmol, 82 mg) and tryptophan methyl ester hydrochloride (0.3 mmol, 76 mg, CAS: 14907-27-8) were dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL), N, N-diisopropyl ethylamine (DIPEA, 0.3 mmol, 39 mg) was added, and after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. TLC was used to detect the progress of the reaction, and the purification method was the same as Example 1-S2 to obtain compound 4i, 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, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated. After standing for 10 h, filtration was carried out to obtain compound 19, a yellow solid 33 mg, with a yield of 28%.

[0133] The reaction occurred in this example is shown in the following formula:

[0134]

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

[0136] NMR data of compound I9, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.28-1.56 (m, 5H), 1.69-1.82 (m, 5H), 2.81 (t, J = 11.4 Hz, 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 = 7.8 Hz, 1H), 7.44 (s, 1H), 7.51 (d, J = 7.8 Hz, 2H), 7.65 (d, J = 8.4 Hz, 1H), 7.73-7.79 (m, 5H), 10.98 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 25.28, 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, 125.43 126.04, 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, found 601.2681.

[0137] Example 10

[0138] A trifluoromethylcyclohexyl benzyl ether substituted amino acid compound is prepared in this example, and the specific structure is shown as formula I10, and the specific process is as follows:

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

[0140] S2. Compound 3a (0.2 mmol, 82 mg) was dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL) with tyrosine methyl ester hydrochloride (0.3 mmol, 70 mg, CAS: 3417-91-2), N, N-diisopropyl ethylamine (DIPEA, 0.3 mmol, 39 mg) was added, after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and stirring was continued. TLC was used to detect the progress of the reaction, and the purification method was the same as that in Example 1-S2 to obtain compound 4j, 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, and stirring was continued at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated. After standing for 10 h, filtration was performed to obtain compound I10, a yellow solid 52 mg, with a yield of 45%.

[0142] The reaction in this example occurred as shown in the following formula:

[0143]

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

[0145] NMR data of compound I10, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.33-1.58 (m, 5H), 1.75-1.87 (m, 5H), 2.89-2.93 (m, 1H), 3.11-3.21 (m, 1H), 3.98 (t, J = 6.6 Hz, 1H), 4.29 (d, J = 2.4 Hz, 2H), 5.26 (s, 2H), 6.74 (d, J = 8.4 Hz, 2H), 7.09 (d, J = 8.4 Hz, 2H), 7.30 (dd, J = 2.4, 9.0 Hz, 1H), 7.44 (1s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.61 (d, 7.8 Hz, 1H), 7.72 (d, J = 7.8 Hz, 1H), 7.77 (s, 1H), 7.84-7.87 (m, 3H); 13 C NMR (150 MHz, MeOD-d 4) δ 26.60, 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 spectral data: HRMS (ESI) calcd for C 34 H 34 F3NO4(M+H) + :578.2513, found 578.2527.

[0146] Example 11

[0147] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure as shown in formula 11, the specific process is:

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

[0149] S2. Compound 3a (0.2 mmol, 82 mg) and methionine methyl ester hydrochloride (0.3 mmol, 60 mg, CAS: 2491-18-1) were dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL), N, N-diisopropyl ethylamine (DIPEA, 0.3 mmol, 39 mg) was added, and after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. TLC was used to detect the progress of the reaction, and the purification method was the same as example 1-S2 to obtain compound 4k, 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, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was added to precipitate, and then the mixture was stirred for 10 h, and then filtered to obtain compound 11, white powder 88 mg, yield 81%.

[0151] The reaction occurred in this example is shown in the following formula:

[0152]

[0153] The test results of the compound obtained in the example are as follows, which show that the compound of formula 111 is indeed obtained.

[0154] NMR data of compound 111, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.34-1.57 (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, 9.0 Hz, 1H), 7.38 (d, J = 1.8 Hz, 1H), 7.55-7.58 (m, 2H), 7.68 (d, J = 8.4 Hz, 1H), 7.74 (s, 1H), 7.84-7.88 (m, 2H), 7.95 (s, 1H); 13 CNMR (150 MHz, MeOD-d 4 ) δ 13.68, 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] A compound of formula 112 is prepared in the example, and the specific structure is shown as formula 112, 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 (0.2 mmol, 82 mg) was dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL) with threonine methyl ester hydrochloride (0.3 mmol, 51 mg, CAS: 62076-66-8), N, N-diisopropyl ethylamine (DIPEA, 0.3 mmol, 39 mg) was added, after stirring at room temperature for 2 h, sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and stirring was continued. TLC was used to detect the progress of the reaction, and the purification method was the same as that in Example 1-S2 to obtain compound 41, a colorless oily solid.

[0159] S3. 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, and stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated, and then stood for 10 h. Filtration obtained compound I12, yellow solid 45 mg, yield 44%.

[0160] The reaction of this example is shown in the following formula:

[0161]

[0162] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which shows that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula I12 is indeed obtained in this example.

[0163] NMR data of compound I12, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.29 (d, J = 6.0 Hz, 3H), 1.36-1.57 (m, 5H), 1.77-1.87 (m, 5H), 2.91 (t, J = 11.4 Hz, 1H), 3.53 (d, J = 7.2 Hz, 1H), 4.08-4.10 (m, 1H), 4.36-4.44 (m, 2H), 5.23 (s, 2H), 7.28 (dd, J = 2.4, 9 Hz, 1H), 7.37 (d, J = 2.4 Hz, 1H), 7.52 (dd, J = 1.2, 8.4 Hz, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.44 (s, 1H), 7.83 (t, J = 9.0 Hz, 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. High-resolution mass spectrometry data: HRMS(ESI) calcd for C 29 H 32 F3NO4(M+H) + :516.2356,found 516.2364.

[0164] Example 13

[0165] This embodiment prepared a trifluoromethylcyclohexylbenzyl ether-substituted aminosulfonic acid compound, the specific structure of which is shown in Formula II1. The specific process is as follows:

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

[0167] S2. Compound 3a (0.2 mmol, 82 mg) and sulphic acid (0.3 mmol, 33 mg, CAS: 13881-91-9) were dissolved in methanol / dichloromethane (V:V = 2 mL: 2 mL). The mixture was stirred at room temperature for 2 h, then sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and stirring continued. The reaction was monitored by TLC. After 3a was completely consumed, the mixture was separated by PTLC (eluent: petroleum ether: ethyl acetate = 2:1) to give compound II1, a yellow solid, 16 mg. Yield: 16%.

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

[0169]

[0170] The test results of the trifluoromethylcyclohexylbenzyl ether-substituted aminosulfonic acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethylcyclohexylbenzyl ether-substituted aminosulfonic acid compounds corresponding to Formula II1.

[0171] NMR data of compound II1 1 H NMR (600MHz, CDCl3-d) 1) δ 1.17-1.42 (m, 6H), 1.69-1.78 (m, 6H), 2.85-2.89 (m, 1H), 4.74 (s, 2H), 5.08 (s, 2H), 7.14-7.16 (m, 2H), 7.37-7.42 (m, 2H), 7.53 (d, J = 7.8 Hz, 1H), 7.65-7.58 (m, 4H). 13 C NMR (150 MHz, CDC13-d 1 ) δ 25.07, 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] Example 14

[0173] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound, the specific structure as shown in formula II2, the specific process is:

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

[0175] S2. Compound 3a (0.2 mmol, 82 mg) and taurine (0.3 mmol, 38 mg, CAS: 107-35-7) were dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL), stirred at room temperature for 2 h, then sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. The purification method is the same as Example 13-S2, and compound II2 is obtained as a colorless oily solid 54 mg. The yield is 52%.

[0176] The reaction occurred in this example is shown in the following formula:

[0177]

[0178] The test results of the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound corresponding to formula II2 is indeed obtained in this example.

[0179] NMR data of compound II2, 1 H NMR (600 MHz, CDC13-d 1) δ 1.18 - 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), 7.15 - 7.17 (m, 2H), 7.38 (dd, J = 1.8, 8.4 Hz, 1H), 7.41 (d, J = 7.8 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.65 - 7.69 (m, 4H); 13 C NMR (150 MHz, CDC13-d 1 ) δ 25.07, 25.78, 33.52, 38.96, 64.52, 68.29, 106.03, 118.19, 122.62, 123.91, 123.95, 124.44, 124.54, 124.94, 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] Example 15

[0181] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound, the specific structure as shown in formula II 3, the specific process is:

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

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

[0184] The reaction of this example is as shown in the following formula:

[0185]

[0186] The test results of the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound corresponding to formula II 3 is indeed obtained in this example.

[0187] NMR data of compound II 3, 1 H NMR (600MHz, CDC13-d1 ) δ 1.25-1.49 (m, 6H), 1.58-1.67 (m, 4H), 1.77-1.86 (m, 6H), 2.93-2.96 (m, 1H), 4.82 (s, 2H), 5.16 (s, 2H), 7.22-7.24 (m, 2H), 7.45 (dd, J = 1.2, 8.4 Hz, 1H), 7.48 (d, J = 7.8 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.72-7.76 (m, 4H); 13 CNMR (150 MHz, CDC13-d 1 ) δ 26.10, 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, 125.57, 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 prepared a trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound, the specific structure as shown in formula II 4, the specific process is:

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

[0191] S2. Compound 3a (0.2 mmol, 82 mg) and 2-aminoethyl hydrogen sulfate (0.3 mmol, 42 mg, CAS: 926-39-6) were dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL), stirred at room temperature for 2 h, then sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. The purification method is the same as Example 13-S2, and compound II 4, yellow solid 10 mg, yield 10% is obtained.

[0192] The reaction occurred in this example is shown in the following formula:

[0193]

[0194] The test results of the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound obtained in this example are as follows, which shows that the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound corresponding to formula II 4 is indeed obtained in this example.

[0195] The nuclear magnetic resonance data of compound II 4,1 H NMR (600 MHz, CDCI3-d 1 ) δ 1.18-1.23 (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), 7.15-7.17 (m, 2H), 7.38 (dd, J = 1.2, 8.4 Hz, 1H), 7.41 (d, J = 8.4 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.65-7.69 (m, 4H); 13 C NMR (150 MHz, CDCI3-d 1 ) δ 26.10, 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] Example 17

[0197] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted aminophosphoric acid compound, the specific structure as shown in formula II 5, the specific process is:

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

[0199] S2. Compound 3a (0.2 mmol, 82 mg) and aminomethyl phosphoric acid (0.3 mmol, 33 mg, CAS: 1066-51-9) were dissolved in methanol / dichloromethane (V:V = 2 mL:2 mL), stirred at room temperature for 2 h, then sodium cyanoborohydride (0.3 mmol, 19 mg) and acetic acid (0.8 mmol, 48 mg) were added, and the stirring was continued. The purification method is the same as Example 13-S2, to obtain compound II 5, brown solid 32 mg, yield 32%.

[0200] The reaction of this example is as shown in the following formula:

[0201]

[0202] The test results of the trifluoromethylcyclohexyl benzyl ether substituted aminophosphoric acid compound obtained in this example are as follows, which shows that the trifluoromethylcyclohexyl benzyl ether substituted aminophosphoric acid compound corresponding to formula II 5 is indeed obtained in this example.

[0203] NMR data of compound II5, 1 H NMR (600 MHz, CDC13-d 1 ) δ 1.26-1.50 (m, 6H), 1.78-1.86 (m, 6H), 2.94 (t, J = 9.6 Hz, 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 = 7.8 Hz, 1H), 7.73-7.76 (m, 4H), 13 C-NMR (150 MHz, CDC13-d 1 ) δ 26.11, 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, 125.57, 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] Example 18

[0205] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted aminophosphoric acid compound, the specific structure as shown in formula II6, the specific process is:

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

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

[0208] The reaction occurred in this example is shown in the following formula:

[0209]

[0210] The test results of the trifluoromethylcyclohexyl benzyl ether substituted aminophosphonic acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted aminophosphonic acid compound corresponding to formula II 6 is indeed obtained in this example.

[0211] NMR data of compound II 6, 1 H NMR (600 MHz, CDC13-d 1 ) δ 1.28-1.47 (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, 8.4 Hz, 1H), 7.48 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.72-7.76 (m, 4H); 13 C NMR (150 MHz, CDC13-d 1 ) δ 26.11, 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, 125.57, 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] Example 19

[0213] A trifluoromethylcyclohexyl benzyl ether substituted aminophosphonic acid compound is prepared in this example, and the specific structure is shown as formula III 1, and the specific process is as follows:

[0214] S1. 3-trifluoromethyl-4-cyclohexyl benzyl alcohol (5.0 mmol, 1290 mg), 4-hydroxybenzaldehyde (5.0 mmol, 611 mg, CAS: 123-08-0) and triphenyl phosphine (PPh3, 6.0 mmol, 1570 mg) were dissolved in anhydrous tetrahydrofuran (10 mL), stirred at room temperature, and diisopropyl azodicarboxylate (DIAD, 6.0 mmol, 1210 mg) was slowly added dropwise within 5 min, and the stirring was continued. The reaction was carried out at room temperature overnight. After the reaction was completed, the reaction solvent was rotary evaporated, and column chromatography was used for separation (eluent was petroleum ether: ethyl acetate = 20: 1) to obtain compound 3b, white powdery solid 996 mg, yield 55%.

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

[0216] S3. Compound 4m obtained in S2 was dissolved in methanol (4 mL), lithium hydroxide aqueous solution (0.5 mol / L, 3.2 mL) was added, stirred at room temperature for 3 h, after the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and then water was added until precipitation was precipitated, and then the mixture was allowed to stand for 10 h, and then filtered to obtain compound III 1, white powder 82 mg, yield 98%.

[0217] The reaction of this example is shown in the following formula:

[0218]

[0219] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula III 1 is indeed obtained in this example.

[0220] NMR data of compound 3b, 1 H NMR (600 MHz, CDCl3-d 1 ) δ 1.28-1.30 (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 = 9.0 Hz, 2H), 7.48 (d, J = 8.4 Hz, 1H), 7.55-7.56 (m, 1H), 7.67 (s, 1H), 7.85 (d, J = 9.0 Hz, 2H), 9.90 (s, 1H); 13 C NMR (150 MHz, CDCl3-d 1) δ 26.08, 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 spectral data: HRMS (ESI) calcd for C 21 H 21 F3O2(M+H) + : 363.1567, found 363.1577.

[0221] Compound III 1 NMR data, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.33-1.56 (m, 5H), 1.76-1.87 (m, 5H), 3.46 (s, 2H), 4.13 (s, 2H), 5.15 (s, 2H), 7.06 (d, J = 8.4 Hz, 2H), 7.41 (d, J = 9.0 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.67 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 25.67, 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 spectral data: HRMS (ESI) calcd for C 23 H 26 F3NO3(M+H) + : 422.1938, found 422.1946.

[0222] Example 20

[0223] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure is shown as formula III 2, the specific process is as follows:

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

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

[0226] S3. Compound 4n obtained in S2 was dissolved in methanol (4 mL), and aqueous lithium hydroxide solution (0.5 mol / L, 3.2 mL) was added, and stirring was continued at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and water was further added until precipitation was precipitated, and the mixture was allowed to stand for 10 h. Filtration obtained compound III2, white powder 24 mg, yield 25%.

[0227] The reaction of this example occurred as shown in the following formula:

[0228]

[0229] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which shows that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula III2 is indeed obtained in this example.

[0230] NMR data of compound III2, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.35-1.56 (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 = 11.4 Hz, 1H), 3.91 (d, J = 15.0 Hz, 1H), 3.96 (dd, J = 3.0, 9.6 Hz, 1H), 4.92 (d, J = 15.0 Hz, 1H), 5.09 (s, 1H), 6.96 (d, J = 8.4 Hz, 2H), 7.15 (d, J = 8.4 Hz, 2H), 7.54 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 1.8 Hz, 1H), 7.67 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4) 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 spectral data: HRMS (ESI) calcd for C 26 H 30 F3NO5(M-H2O+H) + : 476.2043, found 476.2052.

[0231] Example 21

[0232] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure as shown in formula III 3, the specific process is:

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

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

[0235] S3. Compound 4o obtained in S2 was dissolved in methanol (4 mL), lithium hydroxide aqueous solution (0.5 mol / L, 3.2 mL) was added, stirred at room temperature for 3 h, after the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2, and then water was added until precipitation was precipitated, and then the mixture was allowed to stand for 10 h, and then filtered to obtain compound III 3, white powder 16 mg, yield 16%.

[0236] The reaction of this example is shown in the following formula:

[0237]

[0238] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula III 3 is indeed obtained.

[0239] Compound III3NMR data, 1 H NMR (600 MHz, MeOD-d 4 ) δ 1.24-1.47 (m, 5H), 1.66-1.78 (m, 5H), 1.97 (d, J = 3.0 Hz, 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 = 3.0 Hz, 2H), 6.94-6.97 (m, 2H), 7.26-7.33 (m, 2H), 7.45 (d, J = 7.8 Hz, 1H), 7.52 (d, J = 8.4 Hz, 1H), 7.57 (s, 1H); 13 C NMR (150 MHz, MeOD-d 4 ) δ 13.70, 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, found 496.2128.

[0240] Example 22

[0241] This example prepared a trifluoromethylcyclohexyl benzyl ether substituted amino acid compound, the specific structure is shown as formula III4, and the specific process is as follows:

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

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

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

[0245] The reaction in this example occurred as shown in the following formula:

[0246]

[0247] The test results of the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound obtained in this example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted amino acid compound corresponding to formula III 4 is indeed obtained in this example.

[0248] NMR data of compound III 4, 1 H NMR (600 MHz, DMSO-d 6 ) δ 1.15 (d, J = 6.0 Hz, 3H), 1.30-1.56 (m, 5H), 1.71-1.84 (m, 5H), 2.83 (t, J = 11.4 Hz, 1H), 3.09 (t, J = 6.6 Hz, 1H), 3.91-3.95 (m 1H), 4.01 (d, J = 13.2 Hz, 1H), 4.10 (d, J = 13.2 Hz, 1H), 5.16 (s, 2H), 7.04 (d, J = 9.0 Hz, 2H), 7.43 (d, J = 9.0 Hz, 2H), 7.61 (d, J = 7.8 Hz, 1H), 7.67 (d, J = 8.4 Hz, 1H), 7.71 (s, 1H); 13 C NMR (150 MHz, DMSO-d 6 ) δ 20.89, 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, found 466.2205.

[0249] Example 23

[0250] The present example prepared a trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound, the specific structure is shown as formula IV1, and the specific process is as follows:

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

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

[0253] The reaction in the present example is shown in the following formula:

[0254]

[0255] The test results of the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound obtained in the present example are as follows, which show that the trifluoromethylcyclohexyl benzyl ether substituted sulfamic acid compound corresponding to formula IV1 is indeed obtained in the present example.

[0256] NMR data of compound IV1, 1 H NMR (600 MHz, CDCl3-d 1 ) δ 1.27-1.48 (m, 7H), 1.77-1.86 (m, 6H), 1.97 (s, 1H), 2.94 (d, J = 9.6 Hz, 1H), 4.62 (s, 2H), 5.04 (s, 2H), 6.96 (d, J = 9.0 Hz, 2H), 7.29 (d, J = 8.4 Hz, 2H), 7.46 (d, J = 7.8 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H); 13 C NMR (150 MHz, CDCl3-d 1 ) δ 26.10, 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.

[0257] Example 1 HTRF-IP1 method for detecting in vitro agonistic activity of compounds on S1P1 and S1P3

[0258] Cells:

[0259] Chinese hamster ovary-K1 (CHO-K1)Gαqi5 cells (hS1P1-CHO) with stable high S1P1 expression were constructed by Multispan (Hayward, California, USA). 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] Different concentrations of compounds I1-12, II1-6, III1-4, IV1, and the positive control FTY720-P were prepared using 1×Stimb Buffer (containing 1% BSA without free fatty acids).

[0262] instrument:

[0263] EnVision Multifunctional Microplate Reader (PerkinElmer)

[0264] method:

[0265] The concentration of IP1 generated after S1P1 or S1P3 activation was detected using homogeneous time-resolved fluorescence-IP1 (HTRF-IP1) method, and the EC was calculated. 50 The value reflects the receptor-agonistic activity of the compound. IP1 was detected using an IP1 kit (Cisbio). First, hS1P1-CHO and hS1P3-CHO cell buffers were prepared using 1×Stimb Buffer, and 7 μL of a 1×10⁻⁶ concentration was added to each well of a 384-well plate. 7 Cells were added at a concentration of 100 cells / mL. Then, 7 μL of different concentrations of the compound were added to the plate and incubated at 37°C and 5% CO2 for 2 hours. Next, 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 measured using EnVision. The absorbance 665 nm / 615 nm (A) was calculated. 665 / A 615 The ratio of A to B is plotted using nonlinear least squares fitting. 665 / A 615 Standard curve of IP1 concentration versus IP1 concentration. Data statistics:

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

[0267] Experimental results:

[0268] Results are shown in Table 1.

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

[0270]

[0271]

[0272] Experimental conclusion:

[0273] 1) The positive drug FTY720-P has strong agonistic activity on S1P1 and S1P3, EC 50 values are 55.63 nM and 43.27 nM, respectively.

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

[0275] Example 2: Detection of the effect of compounds on rat peripheral lymphocytes

[0276] Animals:

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

[0278] Drugs: FTY720, I6, I11, I12, II3, and II6 were prepared in different concentrations using CMC-Na.

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

[0280] Experimental grouping:

[0281] Normal control group;

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

[0283] Test compound group: 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, 5 rats in each group. The positive drug FTY720 3 mg / kg, 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, II6 3 mg / kg or control solution (0.5% CMC-Na) were administered by gavage. At 0, 2, 4, 8, 12, 24 and 48 hours after administration, blood (20 μL) was collected from the tail vein, and the peripheral blood lymphocytes were counted using a blood cell analyzer (10 9 / L).

[0286] Data statistics:

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

[0288] Experimental results:

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

[0290] Table 2. Maximum percentage reduction of lymphocyte count in rats by compounds

[0291]

[0292]

[0293]

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

[0295] Table 3. Maximum percentage reduction of lymphocyte count in rats by different concentrations of I12

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

[0297] Experimental conclusion:

[0298] 1) The positive drug FTY720 significantly reduced the peripheral blood lymphocytes in rats, P<0.001, with a statistically significant difference.

[0299] 2) Among the test compounds, I12 had the strongest effect on reducing the peripheral blood lymphocytes in rats, and had good dose dependence, P<0.05, with a statistically significant difference.

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

[0301] Animals:

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

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

[0304] Instruments: Mouse lung function instrument (flexiVent TM system, Scireq), electronic balance.

[0305] Experimental grouping:

[0306] Normal control group;

[0307] Model group;

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

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

[0310] Method:

[0311] First, the mice were randomly divided into groups, 14 in the control group and 78 in the modeling group. A single intratracheal instillation of bleomycin 2.5mg / kg was used to construct the mouse pulmonary fibrosis model. After 10 days, the mice after modeling were randomly divided into groups, 16 in the model group and 13 in the drug administration group. The mice in the drug administration group were given daily gavage of 50mg / kg of positive drug nintedanib, 0.3mg / kg of I12, 1mg / kg of I12, and 3mg / kg of I12, and the model group was given control solvents (0.5% CMC-Na). On day 31, the mice were anesthetized and the mouse lung function instrument was used to detect lung function (n=5).

[0312] Data statistics:

[0313] Four indicators of lung function were determined: dynamic compliance Crs, dynamic resistance Rrs, dynamic elasticity Ers, and deep inspiration volume IC. T.TEST was used to compare the data between groups, and P<0.05 was considered to be significantly different.

[0314] Experimental results:

[0315] The results are shown in Figures 3-6 .

[0316] Experimental conclusion:

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

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

[0319] 3) Compound I12 at each concentration significantly improved the lung function of fibrosis mice, and Crs and IC increased significantly, and the Ers index decreased significantly with a certain dose-dependent manner.

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

[0321] Animals:

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

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

[0324] Instruments: ruler, electronic balance.

[0325] Experimental grouping:

[0326] Normal control group;

[0327] Model group;

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

[0329] Method:

[0330] First, the mice were randomly divided into groups, 11 in the control group, 12 in the model group, I12 (0.3 mg / kg), I12 (1 mg / kg), and I12 (3 mg / kg) groups. By giving the mice free access to 2% DSS aqueous solution for 7 days, an acute colitis model in mice was established. On the day of modeling, 0.3 mg / kg of I12, 1 mg / kg of I12, and 3 mg / kg of I12 were administered by gavage, and the model group was given the control solvent (0.5% CMC-Na). After 7 days, the disease activity index DAI of the mice was evaluated, the mice were overdosed, the colon of the mice was taken, the length was measured, and the photograph was taken.

[0331] Data statistics:

[0332] The three indicators of DAI, body weight loss percentage, stool shape, and degree of hematochezia were determined, and the scoring criteria are shown in Table 4:

[0333] Table 4. Criteria for scoring the disease activity index of colitis in mice

[0334]

[0335] Note: ① Normal stool: formed stool; ② Loose stool: paste-like, semi-formed stool that does not adhere to the anus; ③ Diarrhea: watery stool

[0336] The data of each group were compared by T. TEST, and P < 0.05 was considered to be significantly different.

[0337] Experimental results:

[0338] Results are shown in Figures 7-12 .

[0339] Experimental conclusion:

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

[0341] 2) Compound I12 at each concentration significantly improved the colon length of colitis mice, the scores of DAI were significantly decreased compared with the model group, the decrease of body weight was reduced, and the bloody and watery stool was significantly reduced, and it had a certain dose-dependent.

Claims

1. A nitrogen-containing organic acid derivative substituted with trifluoromethylcyclohexylbenzyl ether, characterized in that, Compounds including those with the structure shown in formula (I) and their pharmaceutically acceptable salts: in, R1 is selected from phenyl and naphthyl; R2 is selected from One of them; R3 is selected from H or 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 compounds are selected from the following:

4. A method for synthesizing a nitrogen-containing organic acid derivative substituted with trifluoromethylcyclohexylbenzyl ether as described in any one of claims 1 to 3, characterized in that, Includes the following steps: (1) The product is obtained by photoelectrophoresis of 3-trifluoromethyl-4-cyclohexylbenzyl alcohol as shown in formula (Ⅰa) with 6-OH-2-naphthaldehyde or 4-OH benzaldehyde. (2) The aldehyde group of formula (Ib) reacts with the amino group of NH2R to obtain the trifluoromethylcyclohexylbenzyl ether-substituted nitrogen-containing sulfonic acid / phosphate compound shown in formula (I); (3) The aldehyde group of formula (Ib) reacts with the amino group of NH2R to give the trifluoromethylcyclohexylbenzyl ether-substituted amino acid ester compound shown in formula (Ic); (4) The compound of formula (Ⅰc) is hydrolyzed in an aqueous solution of lithium hydroxide to give the amino acid compound substituted with trifluoromethylcyclohexylbenzyl ether as shown in formula (Ⅰ); 5. The synthesis method according to claim 4, characterized in that, The reaction in step (1) is carried out in tetrahydrofuran or toluene at room temperature, with diisopropyl azodicarbonate and triphenylphosphine added as catalysts. The molar ratio of benzyl alcohol, 6-hydroxy-2-naphthaldehyde or 4-hydroxybenzaldehyde, diisopropyl azodicarbonate 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. After stirring for 2 hours, sodium cyanoborohydride and acetic acid are added as reducing agents, wherein R is selected from alkyl-substituted sulfonic acids and phosphoric acid, and the molar ratio of 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 in step (3) is carried out in methanol / dichloromethane V:V = 1:1 at room temperature, with N,N-diisopropylethylamine added simultaneously. After stirring for 2 hours, sodium cyanoborohydride and acetic acid are added as reducing agents. R is selected from amino acid ester hydrochloride with different substitutions. The molar ratio of 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 in step (4) is carried out in methanol, with the addition of 0.5 mol / L lithium hydroxide aqueous solution, at room temperature, and the molar ratio of the formula (Ⅰc) to lithium hydroxide is 1.0:8.

0.

9. The use of the trifluoromethylcyclohexylbenzyl ether-substituted nitrogen-containing organic acid derivative according to any one of claims 1-3 in the preparation of a medicament for the prevention or treatment of inflammatory diseases.

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

Images