A trifluoromethyl benzyl ether substituted amino acid derivative, its preparation and use

Abstract

The application discloses a trifluoromethyl benzyl ether substituted amino 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 benzyl ether substituted amino acid derivative is obtained through two-step reaction, that is, first, a photo extension reaction is carried out between a benzyl alcohol compound and hydroxynaphthaldehyde (or hydroxybenzaldehyde), then a Schiff base is formed after an amino acid ester hydrochloride, and then the Schiff base is reduced by sodium cyanoborohydride, and then simple hydrolysis is carried out to obtain the trifluoromethyl benzyl ether substituted amino acid derivative. 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 idiopathic pulmonary fibrosis.

Description

Technical Field

[0001] This invention belongs to the field of organic compound synthesis and pharmaceutical technology, specifically relating to a trifluoromethylbenzyl ether-substituted amino acid compound, its synthesis method, and its pharmaceutical uses. Background Technology

[0002] Sphingosine-1 phosphate (S1P) is one of the five G protein-coupled receptor families on the cell membrane surface (S1P). 1-5 Sphingosine-1-phosphate (S1P) is an endogenous ligand that regulates various biological processes, including cell differentiation, vascular stability, inflammation, endothelial integrity, and angiogenesis. S1P is a biologically active sphingolipid involved in the development and progression of many diseases, particularly immune-mediated diseases, and has become an important drug target. Studies have shown that selectively regulating S1P1 without activating S1P3 could be a novel approach for treating idiopathic pulmonary fibrosis (IPF).

[0003] Therefore, exploring the structural characteristics of S1P1, mimicking the binding mode of compounds to receptors, and designing and synthesizing novel selective S1P1 modulators may lead to the discovery of effective drugs for treating IPF. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention designs a series of trifluoromethylbenzyl ether-substituted amino acid compounds. These compounds have a structural feature of a lipophilic trifluoromethylbenzene group as the tail, a naphthalene ring or a benzene ring as the connecting aromatic region, and a carboxylic acid as the hydrophilic polar head. They can also selectively regulate the S1P1 receptor and are expected to be developed into novel agents for the treatment of idiopathic pulmonary fibrosis.

[0005] The present invention also proposes a method for synthesizing the above-mentioned trifluoromethyl benzyl ether-substituted amino acid compounds.

[0006] The present invention also provides a pharmaceutical use of the above-mentioned trifluoromethyl benzyl ether-substituted amino acid compound.

[0007] According to one aspect of the invention, a trifluoromethylbenzyl ether-substituted amino acid compound is provided, comprising compounds with the structure shown in formula (I) and their pharmaceutically acceptable salts:

[0008]

[0009] in,

[0010] R1 is selected from H and trifluoromethyl;

[0011] R2 is selected from phenyl and naphthyl;

[0012] R3 is selected from One of them;

[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 formula (I), when R1 is selected from hydrogen and R2 is selected from a naphthalene ring, R3 is selected from... One of them.

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

[0016] In some embodiments of the present invention, in 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, at least one of the amino acid compounds V1-3, VI1-3, VII1 substituted with trifluoromethylbenzyl ether is present:

[0018]

[0019] According to another aspect of the present invention, a method for synthesizing the trifluoromethylbenzyl ether-substituted amino acid compound is provided, comprising first reacting a benzyl alcohol compound of formula (Ia) with hydroxybenzaldehyde (or hydroxynaphthaldehyde) via photoelectroporation to generate formula (Ib), then reacting it 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 includes at least one of the compounds represented by the following formula:

[0022]

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

[0024]

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

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

[0027] In some embodiments of the present invention, the reaction in 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 in 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 the synthesis formula (Ⅰ) is room temperature (5–30°C).

[0031] In some embodiments of the present invention, the reaction lasts for 12 hours (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 includes purifying the resulting compound after the reaction.

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

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

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

[0037] Beneficial technical effects

[0038] 1. This invention discloses a novel trifluoromethylbenzyl ether-substituted amino acid compound. The synthesis method of this invention is characterized by simple steps, mild reaction conditions, and fast reaction.

[0039] 2. This invention demonstrates, through experimental research on the agonistic activity of trifluoromethylbenzyl ether-substituted amino acid compounds on S1P1, that compounds VI1 and VI2 have high selectivity, strong agonistic activity on S1P1, and no agonistic activity on S1P3. Detailed Implementation

[0040] The present invention will be fully described below with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0041] Example 1

[0042] This embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula V1. The specific process is as follows:

[0043] S1. 3-Trifluoromethylbenzyl alcohol (5.0 mmol, 880 mg, CAS: 349-75-7), 6-hydroxy-2-naphthaldehyde (5.0 mmol, 860 mg), and triphenylphosphine (PPh3, 6.0 mmol, 1570 mg) were dissolved in anhydrous tetrahydrofuran (10 mL). The mixture was stirred at room temperature, and diisopropyl azodicarbonate (DIAD, 6.0 mmol, 1210 mg) was slowly added dropwise over 5 min, with stirring continued. The reaction was carried out overnight at room temperature. After the reaction was complete, the solvent was evaporated, and the mixture was separated by column chromatography (eluent: petroleum ether: ethyl acetate = 20:1) to give compound 3a, a white powder, 924 mg, in a yield of 56%.

[0044] S2. Compound 3a (0.2 mmol, 66 mg) and dimethyl glutamate hydrochloride (0.3 mmol, 64 mg) were dissolved in methanol / dichloromethane (V:V = 2 mL: 2 mL). N,N-diisopropylethylamine (DIPEA, 0.3 mmol, 39 mg) was added, and 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 progress was monitored by TLC. When 3a was completely consumed, saturated sodium bicarbonate solution (20 mL) was added, followed by extraction with ethyl acetate. The organic phase was collected, concentrated under reduced pressure, and separated by PTLC (petroleum ether:ethyl acetate = 2:1) to obtain compound 4a, a colorless oily solid.

[0045] S3. The compound 4a 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 was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound V1, a white powder of 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 trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula V1.

[0049] NMR data of compound 3a 1 H NMR (600MHz, 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 for compound V1 1 H NMR (600MHz, 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 (150MHz, 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 embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula V2. The specific process is as follows:

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

[0054] S2. Compound 3a (0.2 mmol, 66 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, and 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 was continued. The purification method was the same as in Examples 1-S2 to obtain compound 4b, a colorless oily solid.

[0055] S3. The compound 4b obtained in S2 was dissolved in methanol (4 mL), and an aqueous solution of lithium hydroxide (0.5 mol / L, 3.2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound V2, a white powder of 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 trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula V2.

[0059] NMR data for compound V2 1 H NMR (600MHz, 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 (150MHz, 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 embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula V3. The specific process is as follows:

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

[0063] S2. Compound 3a (0.2 mmol, 66 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 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 was continued. The purification method was the same as in Examples 1-S2 to obtain compound 4c, a colorless oily solid.

[0064] S3. The 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 was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound V3, a white powder of 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 trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula V3.

[0068] NMR data for compound V3 1 H NMR (600MHz, 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 (150MHz, DMSO-d) 6δ19.95, 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. High-resolution mass spectrometry data: HRMS(ESI) calcd for C 23 H 22 F3NO4(M+H) + :434.1574,found434.1574.

[0069] Example 4

[0070] This embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula VI1. The specific process is as follows:

[0071] S1. 3,5-Bis(trifluoromethyl)benzyl alcohol (5.0 mmol, 1220 mg, CAS: 32707-89-4), 6-hydroxy-2-naphthaldehyde (5.0 mmol, 860 mg), and triphenylphosphine (PPh3, 6.0 mmol, 1570 mg) were dissolved in anhydrous tetrahydrofuran (10 mL). The mixture was stirred at room temperature, and diisopropyl azodicarbonate (DIAD, 6.0 mmol, 1210 mg) was slowly added dropwise over 5 min, with stirring continued. The reaction was carried out overnight at room temperature. After the reaction was complete, the solvent was evaporated, and the mixture was separated by column chromatography (eluent: petroleum ether: ethyl acetate = 25:1) to give compound 3b, a white powder, 1054 mg, in 53% yield.

[0072] S2. Compound 3b (0.2 mmol, 80 mg) and dimethyl glutamate hydrochloride (0.3 mmol, 64 mg) were dissolved in methanol / dichloromethane (V:V = 2 mL: 2 mL). N,N-diisopropylethylamine (DIPEA, 0.3 mmol, 39 mg) was added, and 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 was continued. The purification method was the same as in Examples 1-S2 to obtain compound 4d, a colorless oily solid.

[0073] S3. The 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 was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound VI1, a white powder of 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 trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula VI1.

[0077] NMR data for compound 3b 1 H NMR (600MHz, 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 δ 68.55, 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 (600MHz, 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 (150MHz, 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 embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula VI2. The specific process is as follows:

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

[0082] S2. Compound 3b (0.2 mmol, 80 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, and 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 was continued. The purification method was the same as in Example 4-S2 to obtain compound 4e, a colorless oily solid.

[0083] S3. The 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 was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound VI2, a white powder of 59 mg, with a yield of 56%.

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

[0085]

[0086] The test results of the trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula VI2.

[0087] NMR data for compound VI2 1 H NMR (600MHz, 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 (150MHz, 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 embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula VI3. The specific process is as follows:

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

[0091] S2. Compound 3d (0.2 mmol, 80 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 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 was continued. The purification method was the same as in Example 4-S2 to obtain compound 4f, a colorless oily solid.

[0092] S3. The 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 was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound VI3, a white powder of 36 mg, with a yield of 36%.

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

[0094]

[0095] The test results of the trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula VI3.

[0096] NMR data for compound VI3 1 H NMR (600MHz, 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 (150MHz, MeOD-d) 4δ19.86, 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 embodiment prepared a trifluoromethylbenzyl ether-substituted amino acid compound, the specific structure of which is shown in Formula VII1. The specific process is as follows:

[0099] S1. 3-Trifluoromethylbenzyl alcohol (5.0 mmol, 880 mg), 4-hydroxybenzaldehyde (5.0 mmol, 611 mg), and triphenylphosphine (PPh3, 6.0 mmol, 1570 mg) were dissolved in anhydrous tetrahydrofuran (10 mL). The mixture was stirred at room temperature, and diisopropyl azodicarbonate (DIAD, 6.0 mmol, 1210 mg) was slowly added dropwise over 5 min, with stirring continued. The reaction was carried out overnight at room temperature. After the reaction was complete, the solvent was evaporated, and the mixture was separated by column chromatography (eluent: petroleum ether: ethyl acetate = 20:1) to give compound 3c, 882 mg of which was a white powder, in 63% yield.

[0100] S2. Compound 3c (0.2 mmol, 56 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 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 was continued. The purification method was the same as in Example 23-S2 to obtain 4 g of the compound 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 was completed, 2 mol / L dilute hydrochloric acid was added to adjust the pH to 1-2. Water was added until a precipitate formed. The mixture was allowed to stand for 10 h and then filtered to obtain compound VII1, a white powder of 62 mg, with a yield of 81%.

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

[0103]

[0104] The test results of the trifluoromethyl benzyl ether-substituted amino acid compounds obtained in this embodiment are as follows, indicating that this embodiment did indeed obtain the trifluoromethyl benzyl ether-substituted amino acid compounds corresponding to Formula VII1.

[0105] 3C NMR data of compound 1 H NMR (600MHz, 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 for compound VII1 1 H NMR (600MHz, 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 (150MHz, MeOD-d) 4δ15.74,15.87,15.99,19.86,49.46,56.06,56.21,65.87,67.58,68.64,115.05,123.54,123.56,123.61,124.22,124.24,129.00,130.71,131.50,138.56,159.35,170.24. 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 for detecting the agonistic activity of compounds on S1P1 and S1P3.

[0109] cell:

[0110] 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.

[0111] drug:

[0112] Different concentrations of compounds V1-3, VI1-3, VII1 and the positive control FTY720-P were prepared using 1×Stimb Buffer (containing 1% BSA without free fatty acids).

[0113] instrument:

[0114] EnVision Multifunctional Microplate Reader (PerkinElmer)

[0115] method:

[0116] 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. 7Cells 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:

[0117] The concentration of the analyte was calculated using a standard curve. EC 50 The values ​​were calculated using Graphpad software 8.0 (Graphpad, LaJolla, CA, USA).

[0118] Experimental results:

[0119] The results are shown in Table 1.

[0120] Table 1. Agonistaltic activity of compounds towards S1P1 and S1P3

[0121]

[0122] Experimental conclusion:

[0123] 1) The positive control drug FTY720-P has strong agonistic activity against S1P1 and S1P3, EC 50 The values ​​are 96.3 nM and 91.2 nM, respectively.

[0124] 2) Compounds VI1 and VI2 have strong agonistic activity towards S1P1, EC 50 The concentrations of 750 nM and 700 nM, respectively, showed no agonistic activity against S1P3.

Claims

1. A trifluoromethylbenzyl ether-substituted amino acid compound, characterized in that, Compounds selected from those with the structure shown in formula (I) and their pharmaceutically acceptable salts: in, R1 is selected from trifluoromethyl; R2 is selected from R3 is selected from One of them.

2. The trifluoromethylbenzyl ether-substituted amino acid compound according to claim 1, characterized in that, The compounds are selected from the following:

3. A method for synthesizing a trifluoromethylbenzyl ether-substituted amino acid compound as described in any one of claims 1 to 2, characterized in that, Includes the following steps: (1) Using benzyl alcohol as a substrate, 6-OH-2-naphthaldehyde was reacted with light to obtain the compound shown in formula (Ib); (2) The aldehyde group of formula (Ib) reacts with the amino group of amino acid ester hydrochloride to obtain the trifluoromethyl benzyl ether-substituted amino acid ester compound shown in formula (Ic). (3) The compound of formula (Ⅰc) is hydrolyzed in an aqueous solution of lithium hydroxide to give the trifluoromethyl benzyl ether-substituted amino acid compound shown in formula (Ⅰ); 4. The synthesis method according to claim 3, characterized in that, The reaction in step (1) is carried out in tetrahydrofuran or toluene at room temperature, which is 5-30°C, with diisopropyl azodicarbonate and triphenylphosphine as catalysts; the molar ratio of benzyl alcohol, 6-hydroxy-2-naphthoaldehyde, diisopropyl azodicarbonate and triphenylphosphine is 1.0:1.0:1.2:1.

2.

5. The synthesis method according to claim 3, characterized in that, The reaction in step (2) is carried out in methanol / dichloromethane V:V = 1:1 at room temperature, which is 5-30℃. N,N-diisopropylethylamine is added at the same time, and after stirring for 2 hours, sodium cyanoborohydride and acetic acid are added as reducing agents. 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.

6. The synthesis method according to claim 3, characterized in that, The reaction in step (3) is carried out in methanol, with the addition of 0.5 mol / L lithium hydroxide aqueous solution, at room temperature (5-30°C), and the molar ratio of formula (I) to lithium hydroxide is 1.0:8.

0.

7. The use of the trifluoromethylbenzyl ether-substituted amino acid compound according to any one of claims 1 to 2 in the preparation of a medicament for the prevention or treatment of inflammation-related diseases, wherein the inflammation-related diseases are selected from pulmonary fibrosis, ulcerative colitis, and multiple sclerosis.

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