Trifluoromethyl benzyl ether substituted amino acid derivative as well as preparation method and application thereof

Abstract

The invention discloses trifluoromethyl benzyl ether substituted amino acid derivatives 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 benzyl ether substituted amino acid compound comprises the following steps: firstly, carrying out a Mitsunobu reaction on a benzyl alcohol compound and hydroxy naphthaldehyde (or hydroxy benzaldehyde), then generating Schiff base with amino-acid ester hydrochloride, then reducing with sodium cyanoborohydride, and then simply hydrolyzing to obtain the trifluoromethyl benzyl ether substituted amino acid compound. 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 idiopathic pulmonary fibrosis.

Description

Technical Field

[0001] The invention belongs to the technical field of organic compound synthesis and medicine, and specifically relates to a trifluoromethyl benzyl ether substituted amino acid compound and a synthesis method 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 approach to treat idiopathic pulmonary fibrosis (IPF).

[0003] Therefore, exploring the structural characteristics of S1P1, simulating the binding mode of compounds with receptors, and targeted design and synthesis of new selective S1P1 modulators are expected to discover effective drugs for the treatment of IPF. Summary of the invention

[0004] In view of the above technical problems, the present invention designs a series of trifluoromethylbenzyl ether substituted amino acid compounds, which have the structural characteristics of a lipophilic group trifluoromethylbenzene as the tail, a naphthalene ring or a benzene ring as the connecting aromatic region, and a carboxylic acid as the hydrophilic polar head. At the same time, they can selectively regulate the S1P1 receptor and are yet to be developed as new preparations for the treatment of idiopathic pulmonary fibrosis.

[0005] The present invention also provides a method for synthesizing the trifluoromethylbenzyl ether substituted amino acid compound.

[0006] The present invention also provides pharmaceutical use of the trifluoromethylbenzyl ether substituted amino acid compound.

[0007] According to one aspect of the present invention, a trifluoromethylbenzyl ether substituted amino acid compound is provided, including a compound of the structure shown in formula (I) and a pharmaceutically acceptable salt thereof:

[0008]

[0009] in,

[0010] R1 is selected from one of H and trifluoromethyl;

[0011] R2 is selected from one of phenyl and naphthyl;

[0012] R3 is selected from One of;

[0013] According to a preferred embodiment of the present invention, at least the following beneficial effects are achieved:

[0014] In some embodiments of the present invention, in the formula (I), when R1 is selected from hydrogen, R2 is selected from naphthalene ring, R3 is selected from One of them.

[0015] In some embodiments of the present invention, in the formula (I), when R1 is selected from hydrogen, R2 is selected from a benzene ring, R3 is selected from

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

[0017] In some embodiments of the present invention, the trifluoromethylbenzyl ether substituted amino acid compounds are at least one of the compounds of the structures shown in V1-3, VI1-3, and VII1:

[0018]

[0019] According to another aspect of the present invention, a method for synthesizing the trifluoromethylbenzyl ether substituted amino acid compound is provided, comprising firstly using a benzyl alcohol compound of formula (Ia) and hydroxybenzaldehyde (or hydroxynaphthaldehyde) to carry out a Mitsunobu reaction to generate formula (Ib), then reacting with an amino acid ester hydrochloride to generate a Schiff base, followed by reduction with sodium cyanoborohydride, and finally hydrolyzing to obtain the trifluoromethylbenzyl ether substituted amino acid compound of formula (I).

[0020]

[0021] In some preferred embodiments of the present invention, the benzyl alcohol compound comprises at least one of the compounds shown in the following formula:

[0022]

[0023] In some preferred embodiments of the present invention, the hydroxybenzaldehyde (or hydroxynaphthaldehyde) 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 step (2) 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 step (2) is room temperature (5-30°C).

[0029] In some embodiments of the present invention, the reaction of step (3) 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 trifluoromethylbenzyl ether substituted amino acid compound, 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 amino acid compounds substituted with trifluoromethyl benzyl ether on S1P1, and proves that compounds VI1 and VI2 have high selectivity, strong agonist activity on S1P1, and no agonist activity on S1P3. DETAILED DESCRIPTION

[0040] 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.

[0041] Example 1

[0042] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula V1, and the specific process is as follows:

[0043] S1. Dissolve 3-trifluoromethylbenzyl alcohol (5.0mmol, 880mg, CAS: 349-75-7), 6-hydroxy-2-naphthaldehyde (5.0mmol, 860mg) 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 3a, a white powdery solid 924mg, with a yield of 56%.

[0044] S2. Compound 3a (0.2mmol, 66mg) 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 continued to stir. TLC was used to monitor the progress of the reaction. When 3a 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 4a as a colorless oily solid.

[0045] S3. Dissolve compound 4a 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 V1 as a white powder (76 mg) with a yield of 83%.

[0046] The reaction that occurs in this embodiment is shown in the following formula:

[0047]

[0048] The test results of the amino acid compound substituted with trifluoromethylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylbenzyl ether corresponding to formula V1 is indeed obtained in this example.

[0049] NMR data of compound 3a, 1 H NMR (600 MHz, CDCl3-d 1 )δ5.26(s,1H),7.26(d,J=2.4Hz,1H),7.32(dd,J=2.4,7.8Hz,1H),7.53(t,J=7.8Hz,1H),7.67(d ,J=7.8Hz,1H),7.77(s,1H),7.80(d,J=8.4Hz,1H),7.92-7.95(m,2H),8.27(s,1H),10.11(s,1H); 13 C NMR (150MHz, CDCl3-d 1 )δ69.36,107.44,12.07,123.91,124.17,124.20,124.22,124.25,125.04,125.07,125.09,125.11,127.87,128.25,129.22,130.68,131.40,132.64,134.2,137.33,138.12,1158.97,191.97. High resolution mass spectrometry data: HRMS (ESI) calcd for C 19 H 13 F3O2(M+H) + :331.0941, found 331.0946.

[0050] NMR data of compound Ⅴ1, 1 H NMR (600 MHz, MeOD-d 4 )δ2.06-2.11(m,1H),2.28-2.35(m,1H),2.41-2.47(m,1H),2.52-2.58(m,1H),4. 00(dd,J=3.0,9.0Hz,1H),4.12(d,J=15.0Hz,1H),5.10(d,J=14.4Hz,1H),5.27(s, 2H),7.23(dd,J=2.4,15.0Hz,1H),7.29(dd,J=1.8,7.8Hz,1H),7.32(d,J=2.4Hz, 1H),7.57(t,J=7.8Hz,1H),7.62(d,J=7.8Hz,2H),7.73-7.77(m,3H),7.80(s,1H);13 C NMR (150 MHz, MeOD-d 4 )δ23.98,30.72,46.76,60.46,70.14,108.39,120.33,124.81,125.07,125.10,125.12,125.15,125.61,125.64,125.66,125.69,126.61,127.79,128.33,128.79,130.43,130.57,131.60,131.81,132.02,132.23,132.43,135.63,140.12,158.18,174.91,178.10. High resolution mass spectrometry data: HRMS (ESI) calcd for C 24 H 22 F3NO5(M-H2O+H) + :444.1417,found444.1427.

[0051] Example 2

[0052] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula V2, and the specific process is as follows:

[0053] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0054] S2. Compound 3a (0.2mmol, 66mg) 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 1-S2 to obtain compound 4b as a colorless oily solid.

[0055] S3. Dissolve compound 4b 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 V2 as a white powder (48 mg) with a yield of 52%.

[0056] The reaction that occurs in this embodiment is shown in the following formula:

[0057]

[0058] The test results of the amino acid compound substituted with trifluoromethylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylbenzyl ether corresponding to formula V2 is indeed obtained in this example.

[0059] NMR data of compound Ⅴ2, 1 H NMR (600 MHz, DMSO-d 6 )δ2.03(s,3H),2.05-2.12(m,2H),2.50-2.67(m,2H),3.77(s,1H),4.18-4.24(m,2H),5.34(s,2H),7.30(dd,J=2.4,9.0Hz,1H), 7.47(d,J=2.4Hz,1H),7.58(dd,J=1.2,8.4Hz,1H),7.65(t,J=7.8Hz,1H),7.72(d,J=7.8Hz,1H),7.83-7.88(m,4H),7.92(s,1H); 13 C NMR (150 MHz, DMSO-d 6 )δ14.31,28.91,29.39,49.68,58.15,68.35,107.21,119.15,123.19,124.07,124.09,124.54,124.57,124.99,126.97,127.77,128.08,128.84,129.00,129.21,129.45,129.53,131.73,133.93,138.28,156.41,170.61. High resolution mass spectrometry data: HRMS (ESI) calcd for C 24 H 24 F3NO3S(M+H) + :464.1502,found464.1503.

[0060] Example 3

[0061] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula V3, and the specific process is as follows:

[0062] S1. The synthesis of compound 3a is the same as that of Example 1-S1.

[0063] S2. Compound 3a (0.2mmol, 66mg) and threonine methyl ester hydrochloride (0.3mmol, 51mg) 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 1-S2 to obtain compound 4c as a colorless oily solid.

[0064] 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) 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 V3 as a white powder (76 mg) with a yield of 88%.

[0065] The reaction that occurs in this embodiment is shown in the following formula:

[0066]

[0067] The test results of the amino acid compound substituted with trifluoromethylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylbenzyl ether corresponding to formula V3 is indeed obtained in this example.

[0068] NMR data of compound Ⅴ3, 1 H NMR (600 MHz, DMSO-d 6 )δ1.11(d,J=6.6Hz,3H),3.03(d,J=5.4Hz,1H),3.88-3.90(m,1H),3.94(d,J =13.2Hz,1H),4.11(d,J=13.2Hz,1H),5.33(s,2H),7.27(dd,J=2.4,9.0Hz,1H ),7.44(d,J=2.4Hz,1H),7.53(dd,J=1.8,8.4Hz,1H),7.65(t,J=7.8Hz,1H), 7.71(d,J=7.8Hz,1H),7.79(d,J=9.0Hz,1H),7.82-7.84(m,3H),7.88(s,1H); 13 C NMR (150 MHz, DMSO-d 6)δ19.95,50.18,65.13,65.64,67.82,106.71,118.35,122.70,123.54,123.56,124.00,124.03,124.50,126.27,127.08,127.24,127.73,128.49,128.70,128.82,129.01,131.20,133.08,137.87,155.58,170.51. HRMS (ESI) calcd for C 23 H 22 F3NO4(M+H) + :434.1574,found434.1574.

[0069] Example 4

[0070] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula VI1, and the specific process is as follows:

[0071] S1. Dissolve 3,5-bis(trifluoromethyl)benzyl alcohol (5.0mmol, 1220mg, CAS: 32707-89-4), 6-hydroxy-2-naphthaldehyde (5.0mmol, 860mg) 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 = 25:1) to obtain compound 3b, 1054mg of white powdery solid, with a yield of 53%.

[0072] S2. Compound 3b (0.2mmol, 80mg) 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 1-S2 to obtain compound 4d as a colorless oily solid.

[0073] 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 VI1 as a white powder (38 mg) with a yield of 36%.

[0074] The reaction that occurs in this embodiment is shown in the following formula:

[0075]

[0076] The test results of the trifluoromethylbenzyl ether substituted amino acid compound obtained in this example are as follows, indicating that the trifluoromethylbenzyl ether substituted amino acid compound corresponding to Formula VI1 is indeed obtained in this example.

[0077] NMR data of compound 3b, 1 H NMR (600 MHz, CDCl3-d 1 )δ5.31(s,2H),7.27(d,J=2.4Hz,1H),7.34(dd,J=2.4,9.0Hz,1H),7.82(d ,J=8.4Hz,1H),7.89(s,1H),7.94-7.97(m,4H),8.29(s,1H),10.11(s,1H); 13 C NMR (150MHz, CDCl3-d 1 )δ68.55,107.45,119.86,122.13,122.16,122.18,122.21,122.23,122.30,123.94,124.11,127.35,127.37,127.92,128.44,131.59,132.00,132.22,132.82,134.17,138.02,138.94,158.54,191.94. High resolution mass spectrometry data: HRMS (ESI) calcd for C 20 H 12 F6O2(M+H) + :399.0814, found 399.0818.

[0078] NMR data of compound VI1, 1 H NMR (600 MHz, DMSO-d 6)δ1.94-1.99(m,1H),2.27-2.32(m,1H),2.34-2.39(m,2H),3.93-3.95(m,1H),4.02(d,J=15.0Hz,1H),4.97(d,J=15.6Hz,1H),5.43( s,2H),7.30-7.32(m,1H),7.44(d,J=2.4Hz,1H),7.66(s,1H),7.78(t,J=8.4Hz,1H),7.84(d,J=9.0Hz,1H),8.10(s,1H),8.23(s,2H); 13 C NMR (150 MHz, DMSO-d 6 )δ22.91,29.53,45.20,58.78,68.15,107.88,119.34,121.08,122.12,122.14,122.89,124.69,126.50,126.80,127.17,127.73,128.84,129.04,129.90,130.51,130.73,130.95,131.16,132.35,133.89,141.05,156.30,173.70,175.00. High resolution mass spectrometry data: HRMS (ESI) calcd for C 25 H 21 F6NO5(M-H2O+H) + :512.1291, found 512.1296.

[0079] Example 5

[0080] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula VI2, and the specific process is as follows:

[0081] S1. The synthesis of compound 3b was the same as that of Example 4-S1.

[0082] S2. Compound 3b (0.2mmol, 80mg) 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 4-S2 to obtain compound 4e as a colorless oily solid.

[0083] 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 VI2 as a white powder (59 mg). The yield was 56%.

[0084] The reaction that occurs in this embodiment is shown in the following formula:

[0085]

[0086] The test results of the trifluoromethylbenzyl ether substituted amino acid compound obtained in this example are as follows, indicating that the trifluoromethylbenzyl ether substituted amino acid compound corresponding to formula VI2 is indeed obtained in this example.

[0087] NMR data of compound VI2, 1 H NMR (600 MHz, Pyridine-d 5 )δ2.05(s,3H),2.16-2.23(m,1H),2.30-2.36(m,1H),2.88-2.91(m,2H),3.81-3.83(m,1H),4.09(d,J=13.2Hz,1H),4.29(d,J=13.2Hz,1H),5 .45(s,2H),7.43(dd,J=2.4,9.0Hz,1H),7.61(d,J=2.4Hz,1H),7.75-7.76(m,1H),7.89-7.92(m,2H),7.99(s,1H),8.10(s,1H),8.23(s,2H); 13 C NMR (150 MHz, Pyridine-d 5 )δ15.05,30.97,33.32,52.23,60.21,68.51,107.96,119.13,123.39,124.83,127.19,127.40,128.18,128.39,129.68,129.99,131.08,131.30,131.52,131.74,134.29,135.45,140.90,149.45,156.50,176.92. High resolution mass spectrometry data: HRMS (ESI) calcd for C 25 H 23 F6NO3S(M+H) + :532.1376,found532.1376.

[0088] Example 6

[0089] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula VI3, and the specific process is as follows:

[0090] S1. The synthesis of compound 3d was the same as that of Example 4-S1.

[0091] S2. Compound 3d (0.2mmol, 80mg) and threonine methyl ester hydrochloride (0.3mmol, 51mg) 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 4-S2 to obtain compound 4f as a colorless oily solid.

[0092] S3. Compound 4f 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 VI3 as a white powder (36 mg). The yield was 36%.

[0093] The reaction that occurs in this embodiment is shown in the following formula:

[0094]

[0095] The test results of the trifluoromethylbenzyl ether substituted amino acid compound obtained in this example are as follows, indicating that the trifluoromethylbenzyl ether substituted amino acid compound corresponding to formula VI3 is indeed obtained in this example.

[0096] NMR data of compound VI3, 1 H NMR (600 MHz, MeOD-d 4 )δ1.26(d,J=0.6Hz,3H),3.22-3.24(m,1H),3.96(t,J=6.6Hz,1H),4.28(d,J=12.6Hz,1H),4.37(d,J=13.2Hz,1H),4.57(s,1H),5 .40(s,1H),7.32(d,J=9.0Hz,1H),7.40(s,1H),7.55(d,J=8.4Hz,1H),7.84(d,J=8.4Hz,2H),7.93(d,J=4.2Hz,2H),8.12(s,2H). 13 C NMR (150 MHz, MeOD-d 4)δ19.86,50.11,65.93,67.77,67.92,107.01,119.18,121.17,122.52,124.33,126.54,127.21,127.40,127.62,129.06,129.62,129.66,131.40,131.62,134.86,140.65,157.02,170.25. High resolution mass spectrometry data: HRMS (ESI) calcd for C 24 H 21 F6NO4(M+H) + :502.1448, found 502.1449.

[0097] Example 7

[0098] This example prepares a trifluoromethyl benzyl ether substituted amino acid compound, the specific structure of which is shown in Formula VII1, and the specific process is as follows:

[0099] S1. Dissolve 3-trifluoromethylbenzyl alcohol (5.0mmol, 880mg), 4-hydroxybenzaldehyde (5.0mmol, 611mg) and triphenylphosphine (PPh3, 6.0mmol, 1570mg) in anhydrous tetrahydrofuran (10m), 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 3c, 882mg of white powdery solid, with a yield of 63%.

[0100] S2. Compound 3c (0.2mmol, 56mg) and threonine methyl ester hydrochloride (0.3mmol, 51mg) 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 23-S2 to obtain compound 4g as a colorless oily solid.

[0101] S3. The colorless oily solid 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 VII as a white powder (62 mg). The yield was 81%.

[0102] The reaction that occurs in this embodiment is shown in the following formula:

[0103]

[0104] The test results of the amino acid compound substituted with trifluoromethylbenzyl ether obtained in this example are as follows, indicating that the amino acid compound substituted with trifluoromethylbenzyl ether corresponding to Formula VII1 is indeed obtained in this example.

[0105] NMR data of compound 3c, 1 H NMR (600 MHz, CDCl3-d 1 )δ5.19(s,2H),7.08(d,J=9.0Hz,2H),7.52-7.55(m,1H),7.62-7.63(m,2H),7.71(s,1H),7.85-7.87(m,2H),9.90(s,1H); 13 CNMR (150MHz, CDCl3-d 1 )δ69.40,115.09,121.24,123.05,124.09,124.12,124.14,124.17,124.85,125.12,125.14,125.17,125.19,126.66,129.25,130.46,130.61,130.85,131.07,131.29,131.50,132.06,137.00,163.28,190.73. High resolution mass spectrometry data: HRMS (ESI) calcd for C 15 H 11 F3O2(M+H) + :281.0784,found281.0786.

[0106] NMR data of compound VII1, 1 H NMR (600 MHz, MeOD-d 4 )δ1.25(d,J=6.0Hz,3H),3.17(d,J=7.8Hz,1H),3.93-3.97(m,1H),4.08(d,J=13.2Hz,1H),4.17(d,J=13.2Hz,1H),7.05( d,J=9.0Hz,2H),7.41(d,J=8.4Hz,2H),7.55(t,J=7.8Hz,1H),7.60(d,J=7.8Hz,1H),7.69(d,J=7.2Hz,1H),7.74(s,1H); 13 C NMR (150 MHz, MeOD-d 4)δ15.74,15.87,15.99,19.86,49.46,56.06,56.21,65.87,67.58,68.64,115.05,123.54,123.56,123.61,124.22,124.24,129.00,130.71,131.50,138.56,159.35,170.24. High resolution mass spectrometry data: HRMS (ESI) calcd for C 19 H 20 F3NO4(M+H) + :384.1417, found 384.1426.

[0107] Pharmacological experiments

[0108] Example 1 In vitro HTRF-IP1 method to detect the agonist activity of compounds on S1P1 and S1P3

[0109] cell:

[0110] 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.

[0111] drug:

[0112] 1×Stimb Buffer (containing 1% BSA without free fatty acids) was used to prepare different concentrations of compounds V1-3, VI1-3, VII1 and the positive drug FTY720-P.

[0113] instrument:

[0114] EnVision Multi-function Microplate Reader (PerkinElmer)

[0115] method:

[0116] 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 7100 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:

[0117] The concentration of the tested compound was calculated based on the standard curve. 50 The values ​​were calculated using Graphpad software 8.0 (Graphpad, La Jolla, CA, USA).

[0118] Experimental results:

[0119] The results are shown in Table 1.

[0120] Table 1. Agonist activity of compounds on S1P1 and S1P3

[0121]

[0122] Experimental conclusion:

[0123] 1) Positive drug FTY720-P has strong agonist activity on S1P1 and S1P3, EC 50 96.3nM and 91.2nM respectively.

[0124] 2) Compounds VI1 and VI2 have strong agonist activity on S1P1, EC 50 The concentrations of the two drugs are 750nM and 700nM respectively, and they have no agonist activity on S1P3.

Claims

1. A trifluoromethylbenzyl ether substituted amino acid compound, characterized in that: Including compounds represented by formula (I) and pharmaceutically acceptable salts thereof: in, R1 is selected from one of H and trifluoromethyl; R2 is selected from one of phenyl and naphthyl; R3 is selected from One of them.

2. The trifluoromethylbenzyl ether substituted amino acid compound according to claim 1, characterized in that: The phenyl and naphthyl groups are: One of them.

3. The trifluoromethylbenzyl ether substituted amino acid compound according to claim 1 or 2, characterized in that: The compound is selected from the following:

4. A method for synthesizing a trifluoromethylbenzyl ether substituted amino acid compound as claimed in any one of claims 1 to 3, characterized in that: The steps include: (1) using benzyl alcohol represented by formula (Ia) as a substrate and performing Mitsunobu reaction with 6-OH, 2-naphthaldehyde or 4-OH benzaldehyde; (2) the aldehyde group of formula (Ib) reacts with the amino group of amino acid ester hydrochloride to obtain a trifluoromethylbenzyl ether substituted amino acid ester compound represented by formula (Ic); (3) The compound of formula (Ic) is hydrolyzed in an aqueous solution of lithium hydroxide to obtain a trifluoromethylbenzyl ether-substituted amino acid compound of 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 of step (2) 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, the reducing agent sodium cyanoborohydride and acetic acid are added. The molar ratio of the formula (Ib), amino acid ester hydrochloride, N,N-diisopropylethylamine, sodium cyanoborohydride and acetic acid is 1.0:1.5:1.5:4.0:1.

0.

7. The synthesis method according to claim 4, characterized in that The reaction in step (3) is carried out in methanol, 0.5 mol / L lithium hydroxide aqueous solution is added, the reaction temperature is room temperature, and the molar ratio of the formula (I) to lithium hydroxide is 1.0:8.

0.

8. Use of the trifluoromethylbenzyl ether substituted amino acid compound according to any one of claims 1 to 3 in the preparation of a drug for preventing or treating inflammation-related diseases.

9. The use according to claim 8, characterized in that The inflammation-related diseases include pulmonary fibrosis, ulcerative colitis, multiple sclerosis, etc.

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