Novel acylphenylalanine derivatives, processes for their preparation and their use as pharmaceuticals

Abstract

The present application relates to a kind of containing effective amount of novel acyl phenylalanine derivative of general formula (I), its preparation method and the use of the derivative as a pharmaceutical composition as a drug, compared with prior art, with more broad application prospect.

Description

Technical Field

[0001] This invention relates to a method for preparing a novel class of acylphenylalanine derivatives and their applications, belonging to the field of medicinal chemistry. The structures of the associated acylphenylalanine derivatives in this invention are unique and novel in this field. Background Technology

[0002] The prevalence of obesity continues to rise worldwide. While rapid modernization, urbanization, and socioeconomic growth have contributed to improved living standards, over the past two decades, high stress levels, sedentary lifestyles, and unhealthy eating habits have led to obesity becoming a global epidemic, affecting almost every organ system and threatening lives. Currently, obesity is one of the most common non-communicable diseases, posing a serious global public health problem.

[0003] The primary cause of obesity is a long-term imbalance between calorie intake and expenditure. Subcutaneous fat is the main storage depot for energy. It is associated with improved metabolism and insulin sensitivity because it comprises brown fat cells containing a large number of mitochondria that play a role in thermogenesis. However, when energy intake is excessive and the storage capacity of subcutaneous fat is depleted, fat accumulates around abdominal visceral organs, such as the liver, leading to non-alcoholic fatty liver disease (NAFLD), atherosclerotic cardiovascular disease (ASCVD) in the vascular system, or insulin resistance in muscles.

[0004] Given these complex connections in the pathogenesis of obesity, current treatment options for managing and treating obesity and type 2 diabetes share some similarities, including lifestyle interventions, drug therapy, various newly developed medical devices, and bariatric surgery, all of which are becoming increasingly popular and technologically advanced. Lifestyle interventions are the foundation and first-line treatment for both obesity and type 2 diabetes, with effective obesity management being paramount. Surgical procedures, whether endoscopic or non-endoscopic, have gained acceptance in patients suitable for different methods, but the unavoidable risks and complications of surgery remind us that these techniques are far from perfect. Therefore, for obese individuals who cannot undergo lifestyle interventions, drug control is a relatively low-risk option, and anti-obesity drugs have some anti-diabetic effects. Currently, the most popular research direction in weight-loss drugs is peptide drugs, such as glucagon-like 1 peptide (GLP-1), which is a hot research topic in the weight-loss field for major pharmaceutical companies. It not only slows gastric emptying, increases satiety, and reduces appetite, but also promotes insulin secretion in a glucose-dependent manner. However, because it is a peptide drug, it can only exert its effects through injection, and there is currently a lack of safe and effective oral weight-loss drugs.

[0005] Recent studies have shown that N-lactic acid-phenylalanine (Lac-Phe), a dipeptide synthesized from lactate and phenylalanine, can inhibit food intake and alleviate obesity. N-lactic acid-phenylalanine is a novel metabolite, and research on it is currently limited. Previous reports have indicated that exercise can increase Lac-Phe levels, and that the in vitro cytosolic enzyme CNDP2 can catalyze Lac-Phe synthesis through the condensation of lactate and phenylalanine. Studies have found that injecting Lac-Phe into obese mice fed a high-fat diet significantly inhibited food intake, reduced obesity, and improved glucose homeostasis (Nature, 2022, 606, 785-790). However, these studies are limited to injectable administration, and oral administration is ineffective. Therefore, developing orally administered Lac-Phe derivatives is highly promising and could potentially change the current situation of a lack of safe and effective oral weight-loss drugs on the market. Furthermore, in-depth research into other pharmaceutical uses of these derivatives is expected to expand their indications and lay the foundation for the development of safer and more effective drugs or health products. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide a novel method for preparing acylphenylalanine derivatives for the first time, which provides possibilities for the treatment and prevention of diseases such as obesity, hyperlipidemia, non-alcoholic fatty liver, drug-induced liver injury, and diabetes.

[0007] The novel acylphenylalanine derivative of this invention is a compound containing an effective amount of the general formula (I):

[0008]

[0009] Wherein, R1 is selected from the following structures:

[0010] R can be selected from hydrogen, alkyl, alkoxy, halogen, cycloalkyl, or trifluoromethyl.

[0011] R2 is selected from the following structures:

[0012]

[0013] Preferably, the compound having general formula (I) as defined in this invention:

[0014] Wherein, R1 is selected from the following structures:

[0015] R can be selected from hydrogen, C1-C6 alkyl, alkoxy, chlorine, fluorine, C3-C6 cycloalkyl, or trifluoromethyl.

[0016] R2 is selected from the following structures:

[0017]

[0018] More preferred compounds of the present invention include, but are not limited to:

[0019] ((S)-2-hydroxybutyryl)-L-phenylalanine (4),

[0020] ((S)-2-hydroxy-3-phenylpropionyl)-L-phenylalanine (7),

[0021] ((S)-2-hydroxypropionyl)-D-phenylalanine (8),

[0022] (S)-3-(4-fluorophenyl)-2-((S)-2-hydroxy-3-methylbutyramide)propionic acid (9),

[0023] (S)-3-(4-(benzyloxy)phenyl)2-propamidopropionic acid (13),

[0024] (S)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (14),

[0025] (R)-3-(4-(benzyloxy)phenyl)-2-((R)-2-hydroxy-3-methylbutyramido)propionic acid (15),

[0026] (R)-3-(4-(benzyloxy)phenyl)-2-propamidopropionic acid (16),

[0027] (R)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (17),

[0028] (R)-3-(4-(benzyloxy)phenyl)-2-(cyclopropaneformamido)propionic acid (18),

[0029] (R)-3-(4-((4-fluorobenzyl)oxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (19),

[0030] (R)-3-(4-(cyclopropylmethoxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (20),

[0031] (R)-2-((S)-2-hydroxypropamido)-3-(4-methoxyphenyl)propionic acid (21).

[0032] Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective dose of the compound and a suitable carrier, diluent or excipient.

[0033] This invention also relates to the use of the said compound or pharmaceutical composition thereof in the preparation of a medicament for the prevention and / or treatment of at least one of the following diseases: obesity, hyperlipidemia, non-alcoholic fatty liver disease, alcohol poisoning, alcoholic steatohepatitis, drug-induced liver injury, dyslipidemia, organ fibrosis, cirrhosis, liver failure, diabetes, diabetic complications, hyperuricemia, gout, altitude sickness, cerebral hypoxia, cerebral ischemia, and myocardial infarction.

[0034] Detailed description of the invention

[0035] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0036] Any general formula or structure shown herein, including compounds of general formula (I), is also intended to represent both unlabeled and isotopically labeled forms of said compounds. Examples of isotopes that may be contained in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine. In the compounds of the present invention, any atom not explicitly designated as a particular isotope is intended to represent any stable isotope of that atom. Unless otherwise stated, when a position is explicitly designated as “H” or “hydrogen”, it should be understood that the position is composed of hydrogen in its isotopic composition. Therefore, in the compounds of the present invention, any atom explicitly designated as deuterium (D) is intended to represent deuterium.

[0037] "Pharmaceutical composition" refers to a mixture containing one or more of the compounds described in this invention, along with other chemical components such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to promote the absorption of the active ingredient by an organism, thereby facilitating its biological activity within the body. Attached Figure Description

[0038] Figure 1 Compound 18 improved the histological characteristics of DIO mice: (A) mouse body weight; (B) mouse liver weight; (C) mouse white adipose tissue weight; (D) HE-stained liver sections, scale bar: 50 μm. *P<0.05, **P<0.01, ***P<0.001, compared with the model group.

[0039] Figure 2 Effects of compound 18 on lipid metabolism in DIO mice: (A) liver TC level; (B) liver TG level; (C) serum TG level. One-way ANOVA and Tukey's multiple-comparison post-hoc test were used for analysis (n=6). *P<0.05, **P<0.01, ***P<0.001, compared with the model group.

[0040] Figure 3Compound 18 improved the histological characteristics of NASH mice: (A) NAS score; (B) HE-stained liver sections, scale bar: 100 μm. Analysis was performed using one-way ANOVA and Tukey's multiple-comparison post-hoc test (n=6). *P<0.05, **P<0.01, ***P<0.001, compared with the model group.

[0041] Figure 4 Effects of compound 18 on lipid metabolism and hepatotoxicity in NASH mice: (A) hepatic TC level; (B) hepatic TG level; (C) serum TG level; (D) serum ALT level; (E) serum AST level. One-way ANOVA and Tukey's multiple-comparison post-hoc test were used for analysis (n=6). *P<0.05, **P<0.01, ***P<0.001, compared with the model group.

[0042] Figure 5 Effects of compounds 1, 18, and 19 on hepatotoxicity in mice with APAP-induced liver injury: (A) serum AST level; (B) serum ALT level; (C) HE-stained liver sections. Statistical analysis was performed using one-way ANOVA and Tukey's multiple-comparison post-hoc test (n=6). *P<0.05, **P<0.01, ***P<0.001, compared with the model group. Detailed Implementation

[0043] The present invention will be further described below with reference to embodiments. It should be noted that the following embodiments are for illustrative purposes only and are not intended to limit the present invention. Various modifications made by those skilled in the art based on the teachings of the present invention should be within the scope of protection of the claims of this application.

[0044] Synthesis and preparation of the compounds of this invention:

[0045] Synthesis Route 1:

[0046]

[0047] Reagents and conditions: (i) ia: HBTU, DIPEA, DCM, rt, 10 h, 60%; ib: pyridine, DCM, 0℃, 3 h, 80%; (ii) LiOH·H2O, HF / MeOH / H2O, room temperature, 4 h, 98%.

[0048] (ia): HBTU (1.2 equiv), DIPEA (3 equiv), and the desired amine (1.2 equivalents) were added to a dichloromethane (10 mL) solution of the selected carboxylic acid. After stirring at room temperature for 12 hours, the reaction mixture was concentrated, washed successively with saturated sodium bicarbonate solution and 1N HCl, and extracted with ethyl acetate. The product was then purified by silica gel column chromatography using petroleum / ethyl acetate (3:1; v / v) to give the product.

[0049] (ib): At 0°C, a solution of acyl chloride (1 equiv) in dichloromethane (5 mL) was slowly added to a stirred solution of phenylalanine methyl ester (1.2 equiv) in dichloromethane (5 mL) and pyridine (0.5 mL). After stirring for another 3 hours at room temperature, 4 mL of water was added to quench the reaction. The organic layer was concentrated under vacuum and the mixture was diluted with water. The aqueous layer was adjusted to pH 4–5 with 1 N HCl and extracted with ethyl acetate. The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, concentrated under vacuum, and then purified by silica gel column chromatography using petroleum / ethyl acetate (3:1; v / v) to give the intermediate.

[0050] (ii): The obtained intermediate was dissolved in a 3:3:1 THF / MeOH / H2O (7 mL) solution, and LiOH·H2O (3 equiv) was added. After stirring at room temperature for 4 hours, the solution was acidified with hydrochloric acid, filtered, and the filter cake was washed with cold water to obtain the compound.

[0051] Synthesis Route 2:

[0052]

[0053] Reagents and conditions: (i) K2CO3, MeCN, reflux, 3 h, yield 87-89%; (ii) TFA, DCM, 0℃, 3 h, yield 88%; (iii) ① HBTU, DIPEA, DCM, rt, 12 h, 60%; ② LiOH·H2O, THF / MeOH / H2O, room temperature, 4 h, 98%. (i): Potassium carbonate (3.00 equiv) and ethyl 8-bromooctanoate (1.10 equiv) were added to 10 mL of MeCN containing butoxycarbonyl-D-tyrosine-methoxy ester (1.00 equiv), and the mixture was stirred at 70℃ for 3 h. After the reaction was complete, the mixture was cooled to room temperature, filtered under reduced pressure, and the filtrate was evaporated to dryness. The filtrate was then purified by silica gel column chromatography using petroleum / ethyl acetate (5:1; v / v) to obtain the intermediate product.

[0054] (ii): At 0°C, TFA (3 equiv) was slowly added dropwise to a 10 mL solution of the selected amide in dichloromethane. After stirring at room temperature for 4 hours, the reaction mixture was concentrated and extracted with saturated sodium bicarbonate solution and ethyl acetate (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give the product.

[0055] (iii): HBTU (1.2 equiv), DIPEA (3 equiv), and the desired amine (1.2 equiv) were added to a dichloromethane (10 mL) solution of the selected carboxylic acid. After stirring at room temperature for 16 hours, the reaction mixture was concentrated, washed successively with saturated sodium bicarbonate solution and 1N HCl, and extracted with ethyl acetate. The intermediate was then purified by silica gel column chromatography using petroleum / ethyl acetate (5:1; v / v). The obtained intermediate was dissolved in a 3:3:1 THF / MeOH / H2O (7 mL) solution, and LiOH·H2O (3 equiv) was added. After stirring at room temperature for 3 hours, the solution was acidified with hydrochloric acid, filtered, and the filter cake was washed with cold water to obtain the compound.

[0056] Example 1

[0057] ((S)-2-hydroxypropionyl)-L-phenylalanine(1)

[0058]

[0059] (S)-2-amino-3-phenylpropionate methyl hydrochloride (1.0 equiv), sodium propionate (1.2 equiv), HBTU (1.2 equiv), and DIPEA (3 equiv) were dissolved in dichloromethane and stirred at room temperature for 10 h. After the reaction was complete, the reaction mixture was concentrated, washed successively with saturated sodium bicarbonate solution and 1N HCl, and extracted with ethyl acetate. The mixture was then purified by silica gel column chromatography using petroleum / ethyl acetate (3:1; v / v) to obtain the target compound. Yield: 51%, white powder. 1 H NMR (400MHz, DMSO-d6) δ7.61 (d, J=8.2Hz, 1H), 7.30-7.24 (m, 2H), 7.23-7.19 (m, 1H), 7.19-7.15 (m, 2H), 4.51 (tdJ=8.0 , 5.2Hz, 1H), 3.93 (q, J=6.7Hz, 1H), 3.10 (dd, J=13.7, 5.1Hz, 1H), 3.01 (dd, J=13.7, 8.0Hz, 1H), 1.10 (d, J=6.8Hz, 3H). 13C NMR (101MHz, DMSO-d6) δ174.17, 172.74, 137.20, 129.28, 128.17, 126.52, 67.11, 52.36, 36.62, 21.02.ESI-MS m / z: [MH] - calcd.for C 12 H 14 NO4 - , 236.0928; Found, 236.0920.

[0060] Acetyl-L-phenylalanine (2)

[0061]

[0062] Synthetic method is the same as compound 1. Yield: 88%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.64 (s, 1H), 8.18 (d, J=8.1Hz, 1H), 7.31-7.25 (m, 2H), 7.25-7.17 (m, 3H), 4 .40 (td, J=9.1, 5.0Hz, 1H), 3.04 (dd, J=13.8, 4.9Hz, 1H), 2.83 (dd, J=13.8, 9.6Hz, 1H), 1.78 (s, 3H). 13 CNMR (101MHz, DMSO-d6) δ173.20, 169.26, 137.75, 129.07, 128.20, 126.42, 53.52, 36.79, 22.35.ESI-MS m / z: [MH] - calcd.for C 11 H 12 NO3 - , 206.0823; Found, 206.0821.

[0063] Propionyl-L-phenylalanine (3)

[0064]

[0065] Synthetic method is the same as compound 1. Yield: 62%, white powder. 11H NMR (400MHz, DMSO-d6) δ12.63 (s, 1H), 8.08 (d, J = 8.1Hz, 1H), 7.30-7.24 (m, 2H), 7.24-7.16 (m, 3H), 4.41 (td, J = 9.1, 4.9Hz, 1H), 3.05 (dd, J=13.8, 4.8Hz, 1H), 2.84 (dd, J=13.7, 9.7Hz, 1H), 2.05 (q, J=7.5Hz, 2H), 0.90 (t, J=7.6Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.25, 172.93, 137.77, 129.10, 128.14, 126.38, 53.33, 36.80, 28.22, 9.79.ESI-MS m / z: [MH] - calcd.forC 12 H 14 NO3 - , 220.0979; Found, 220.0971.

[0066] ((S)-2-hydroxybutyryl)-L-phenylalanine (4)

[0067]

[0068] Synthetic method is the same as compound 1. Yield: 58%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.8.2 (s, 1H), 7.63 (d, J=8.2Hz, 1H), 7.30-7.23 (m, 2H), 7.22-7.14 (m, 3H), 5.51 (s, 1H), 4.54 (td, J=8.2, 5.1Hz, 1H), 3.78 (s, 1H), 3.09 (dd, J=13.8, 4.9Hz, 1H), 3.00 (dd, J=13.8, 8.2Hz, 1H), 1.59-1.49 (m, 1H), 1.43-1.33 (m, 1H), 0.72 (t, J=7.4Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.44, 172.79, 137.26, 129.20, 128.16, 126.48, 71.80, 52.39, 36.70, 27.31, 9.14.ESI-MS m / z: [MH] - calcd.for C 13 H 16 NO4 -, 250.1085; Found, 250.1081.

[0069] ((S)-2-hydroxy-3-methylbutyryl)-L-phenylalanine (5)

[0070]

[0071] Synthetic method is the same as compound 1. Yield: 55%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.78 (s, 1H), 7.65 (d, J=8.3Hz, 1H), 7.28-7.23 (m, 2H), 7.22-7.15 (m, 3H), 4.56 (td, J=8.4, 5.1Hz, 1H), 3.65 (d, J =3.7Hz, 1H), 3.08 (dd, J=13.9, 5.0Hz, 1H), 3.00 (dd, J=13.9, 8.6Hz, 1H), 1.94-1.80 (m, 1H), 0.79 (d, J=6.9Hz, 3H), 0.57 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.08, 172.84, 137.34, 129.15, 128.19, 126.46, 75.07, 52.43, 36.80, 31.23, 19.12, 15.94.ESI-MS m / z: [MH] calcd.for C 14 H 18 NO4 - , 264.1241; Found, 264.1238.

[0072] ((S)-2-hydroxy-2-phenylacetyl)-L-phenylalanine (6)

[0073]

[0074] Synthetic method is the same as compound 1. Yield: 55%, white powder. 1H NMR (400MHz, DMSO-d6) δ12.87 (s, 1H), 8.06 (d, J=8.1Hz, 1H), 7.38-7.30 (m, 2H), 7.30-7.21 (m, 5H), 7.20-7.13 (m, 2H), 7.13-7.07 (m, 1H), 6.22-6.08 (m, 1H), 4.93 (s, 1H), 4.47 (td, J=8.4, 4.2Hz, 1H), 3.09 (dd, J=13.8, 4.3Hz, 1H), 3.01 (dd, J=13.6, 8.4Hz, 1H). 13 C NMR (101MHz, DMSO-d6) δ172.66, 171.83, 141.15, 137.31, 129.19, 128.21, 127.85, 127.34, 126.80, 126.47, 73.05, 52.90, 36.50.ESI-MS m / z: [MH] - calcd.for C 17 H 16 NO4 - , 298.1085; Found, 298.1081.

[0075] ((S)-2-hydroxy-3-phenylpropionyl)-L-phenylalanine (7)

[0076]

[0077] Synthetic method is the same as compound 1. Yield: 61%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.92 (s, 1H), 7.66 (d, J=8.2Hz, 1H), 7.30-7.22 (m, 4H), 7.22-7.17 (m, 4H), 7.06 (d, J=6.8Hz, 2H), 5.72 (d, J=5.3H z, 1H), 4.55 (td, J=8.2, 3.3Hz, 1H), 4.12-4.03 (m, 1H), 2.99 (d, J=6.3Hz, 2H), 2.88 (dd, J=13.8, 3.4Hz, 1H), 2.61 (dd, J=13.8, 8.3Hz, 1H). 13 C NMR (101MHz, DMSO-d6) δ173.34, 173.05, 138.84, 137.46, 130.00, 129.70, 128.62, 128.39, 126.96, 126.51, 72.39, 52.81, 40.79, 37.23.ESI-MS m / z:[MH] -calcd.for C 18 H 18 NO4 - , 312.1241; Found.312.1237.

[0078] ((S)-2-hydroxypropionyl)-D-phenylalanine (8)

[0079]

[0080] Synthetic method is the same as compound 1. Yield: 56%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.75 (s, 1H), 7.68 (d, J=8.2Hz, 1H), 7.30-7.23 (m, 2H), 7.23-7.15 (m, 3H), 5.49 (s, 1H), 4.47 (td, J =8.2, 5.0Hz, 1H), 3.94 (q, J=6.7Hz, 1H), 3.08 (dd, J=13.8, 5.0Hz, 1H), 3.00 (dd, J=13.8, 8.3Hz, 1H), 1.13 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ174.25, 172.73, 137.35, 129.19, 128.21, 126.50, 66.94, 52.66, 36.58, 20.96.ESI-MS m / z: [MH] - calcd.for C 12 H 14 NO4 - , 236.0928; Found, 236.0924.

[0081] (S)-3-(4-fluorophenyl)-2-((S)-2-hydroxy-3-methylbutyramide)propionic acid (9)

[0082]

[0083] Synthetic method is the same as compound 1. Yield: 55%, white powder. 1H NMR (400MHz, DMSO-d6) δ12.86 (s, 1H), 7.70 (d, J=8.4Hz, 1H), 7.29-7.20 (m, 2H), 7.07 (t, J=8.8Hz, 2H), 5.45 (s, 1H), 4.54 (td, J=8.7, 5.0Hz, 1H), 3 .64 (d, J=3.4Hz, 1H), 3.07 (dd, J=13.8, 4.8Hz, 1H), 2.99 (dd, J=14.0, 8.9Hz, 1H), 1.94-1.78 (m, 1H), 0.78 (d, J=6.9Hz, 3H), 0.54 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6)) δ173.10, 172.79, 161.08 (d, J=243.41Hz), 133.61 (d, J=3.03Hz), 130 .98 (d, J=8.08Hz), 114.85 (d, J=21.21Hz), 75.10, 52.44, 35.86, 31.23, 19.06, 15.91.ESI-MS m / z:[MH] - calcd.for C 14 H 17 FNO4 - , 282.1147; Found, 282.1143.

[0084] Propionyl-D-phenylalanine (10)

[0085]

[0086] Synthetic method is the same as compound 1. Yield: 59%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.66 (s, 1H), 8.09 (d, J=8.1Hz, 1H), 7.31-7.24 (m, 2H), 7.24-7.15 (m, 3H), 4.41 (td, J=9.4, 4.9Hz, 1H), 3.04 (dd, J=13.7, 4.8Hz, 1H), 2.84 (dd, J=13.7, 9.8Hz, 1H), 2.05 (q, J=7.4Hz, 2H), 0.90 (t, J=7.6Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.25, 172.93, 137.78, 129.10, 128.14, 126.38, 53.34, 36.81, 28.22, 9.79.ESI-MS m / z: [MH] -calcd.forC 12 H 14 NO3 - , 220.0979; Found, 220.0972.

[0087] ((S)-2-hydroxy-3-methylbutyryl)-D-phenylalanine (11)

[0088]

[0089] Synthetic method is the same as compound 1. Yield: 58%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.78 (s, 1H), 7.73 (d, J=8.2Hz, 1H), 7.30-7.24 (m, 2H), 7.23-7.15 (m, 3H), 5.33 (s, 1H), 4.50 (td, J=8.4, 4.9Hz, 1H ), 3.65 (s, 1H), 3.09 (dd, J=13.8, 4.9Hz, 1H), 2.99 (dd, J=13.9, 8.6Hz, 1H), 1.95-1.86 (m, 1H), 0.83 (d, J=6.9Hz, 3H), 0.67 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.27, 172.82, 137.43, 129.15, 128.24, 126.48, 74.97, 52.69, 36.63, 31.35, 19.21, 15.87.ESI-MS m / z: [MH]-calcd.for C 14 H 18 NO4 - , 264.1241; Found, 264.1236.

[0090] (cyclopropane carbonyl)-D-phenylalanine (12)

[0091]

[0092] Synthetic method is the same as compound 1. Yield: 67%, white powder. 1H NMR (400MHz, DMSO-d6) δ12.64 (s, 1H), 8.40 (d, J=8.0Hz, 1H), 7.32-7.25 (m, 2H), 7.25-7.16 (m, 3H), 4.42 (dt, J=9 .2, 5.0Hz, 1H), 3.04 (dd, J=13.8, 5.0Hz, 1H), 2.86 (dd, J=13.8, 9.5Hz, 1H), 1.66-1.54 (m, 1H), 0.69-0.49 (m, 4H). 13 C NMR (101MHz, DMSO-d6) δ173.26, 172.74, 137.73, 129.10, 128.21, 126.43, 53.64, 36.91, 13.35, 6.46, 6.35.ESI-MS m / z: [MH] - calcd.forC 13 H 14 NO3 - , 232.0979; Found, 232.0971.

[0093] (S)-3-(4-(benzyloxy)phenyl)-2-propamidopropionic acid (13)

[0094]

[0095] Synthetic method is the same as compound 1. Yield: 54%, white powder. 1H NMR (400MHz, DMSO-d6) δ 12.63 (s, 1H), 8.05 (d, J = 8.1Hz, 1H), 7.43 (d, J = 7.1Hz, 2H), 7.38 (t, J = 7.3Hz, 2H), 7.32 (t, J = 7.1Hz, 1H), 7.14 (d, J = 8.5Hz, 2H), 6.91 (d, J = 8.5Hz, 2H). J=8.6Hz, 2H), 5.05 (s, 2H), 4.36 (td, J=9.2, 5.0Hz, 1H), 2.97 (dd, J=13.8, 4.8Hz , 1H), 2.77 (dd, J=13.8, 9.6Hz, 1H), 2.06 (q, J=7.5Hz, 2H), 0.91 (t, J=7.6Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.33, 172.92, 157.00, 137.21, 130.15, 129.86, 128.43, 127.80, 127.69, 114.45, 69.12, 53.59, 35.98, 28.22, 9.82.ESI-MS m / z:[MH] -calcd.for C 19 H 20 NO4 - , 326.1398; Found, 326.1394.

[0096] (S)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (14)

[0097]

[0098] Synthetic method is the same as compound 1. Yield: 50%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.88 (s, 1H), 7.58 (d, J=8.1Hz, 1H), 7.47-7.41 (m, 2H), 7. 41-7.35 (m, 2H), 7.35-7.29 (m, 1H), 7.09 (d, J = 8.6Hz, 2H), 6.91 (d, J = 8.7Hz, 2H), 5 .72-5.51 (m, 1H), 5.05 (s, 2H), 4.45 (td, J=7.8, 5.2Hz, 1H), 3.94 (q, J=6.7Hz, 1H), 3.03 (dd, J=13.8, 5.1Hz, 1H), 2.94 (dd, J=13.8, 7.7Hz, 1H), 1.12 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ174.12, 172.77, 157.11, 137.18, 130.29, 129.21, 1 28.42, 127.80, 127.69, 114.47, 69.14, 67.11, 52.55, 35.78, 21.04.ESI-MS m / z:[MH] - calcd.for C 19 H 20 NO5 - , 342.1347; Found, 342.1343.

[0099]

[0100] (R)-3-(4-(benzyloxy)phenyl)-2-((R)-2-hydroxy-3-methylbutyramido)propionic acid (15) was synthesized by the same method as compound 1. Yield: 58%, white powder. 1H NMR (400MHz, DMSO-d6) δ12.76 (s, 1H), 7.61 (d, J=8.3Hz, 1H), 7.43 (d, J=7.0Hz, 2H), 7.38 (t, J=7 .3Hz, 2H), 7.31 (t, J=7.1Hz, 1H), 7.11 (d, J=8.6Hz, 2H), 6.90 (d, J=8.6Hz, 2H), 5.45 (d, J=5.6Hz, 1H), 5.05 (s, 2H), 4.51 (td, J=8.1, 5.3Hz, 1H), 3.66 (t, J=4.3Hz, 1H), 3.00 (dd, J=14.1, 5.3Hz, 1H ), 2.94 (dd, J=14.0, 8.2Hz, 1H), 1.93-1.83 (m, 1H), 0.81 (d, J=6.9Hz, 3H), 0.60 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.48, 173.33, 157.55, 137.66, 130.63, 129.80, 128.87, 128.22, 128.08, 114.97, 75.52, 69.59, 53.03, 36.48, 31.70, 19.58, 16.42.ESI-MS m / z:[MH] - calcd.for C 21 H 24 NO5 - , 370.1660; Found, 370.1655.

[0101] (R)-3-(4-(benzyloxy)phenyl)-2-propamidopropionic acid (16)

[0102]

[0103] Potassium carbonate (3.00 equiv) and ethyl 8-bromooctanoate (1.10 equiv) were added to 10 mL of MeCN containing butoxycarbonyl-D-tyrosine-methoxy ester (1.00 equiv), and the mixture was stirred at 70 °C for 3 hours. After the reaction was complete, the mixture was cooled to room temperature, filtered under reduced pressure, and the filtrate was evaporated to dryness. The filtrate was then purified by silica gel column chromatography using petroleum ether / ethyl acetate (5:1; v / v) to give intermediate 16a. TFA (3 equiv) was slowly added dropwise to a 10 mL solution of 16a in dichloromethane at 0 °C. After stirring at room temperature for 4 hours, the reaction mixture was concentrated and extracted with saturated sodium bicarbonate solution and ethyl acetate (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give intermediate 16b. HBTU (1.2 equiv), DIPEA (3 equiv), and propionic acid (1.2 equiv) were added to a 10 mL solution of 16b in dichloromethane. After stirring at room temperature for 16 hours, the reaction mixture was concentrated, washed successively with saturated sodium bicarbonate solution and 1N HCl, and extracted with ethyl acetate. The product was then purified by silica gel column chromatography using petroleum / ethyl acetate (10:7; v / v) to give product 16. Yield: 59%, white powder. 1H NMR (400MHz, DMSO-d6) δ 12.62 (s, 1H), 8.05 (d, J = 8.1Hz, 1H), 7.43 (d, J = 7.0Hz, 2H), 7.38 (t, J = 7.3Hz, 2H), 7.32 (t, J = 7.1Hz, 1H), 7.14 (d, J = 8.6Hz, 2H), 6.91 (d, J = 8.6Hz, 2H). J=8.6Hz, 2H), 5.05 (s, 2H), 4.36 (td, J=9.2, 4.9Hz, 1H), 2.97 (dd, J=13.8, 4.9Hz , 1H), 2.78 (dd, J=13.8, 9.6Hz, 1H), 2.06 (q, J=7.6Hz, 2H), 0.91 (t, J=7.6Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ173.33, 172.92, 157.00, 137.21, 130.15, 129.87, 128.43, 127.80, 127.69, 114.45, 69.12, 53.60, 35.99, 28.22, 9.82.ESI-MS m / z:[MH] - calcd.for C 19 H 20 NO4 - , 326.1398; Found, 326.1391.

[0104] (R)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (17)

[0105]

[0106] Synthetic method is the same as compound 16. Yield: 56%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.88 (s, 1H), 7.58 (d, J=8.1Hz, 1H), 7.47-7.41 (m, 2H), 7. 41-7.35 (m, 2H), 7.35-7.29 (m, 1H), 7.09 (d, J = 8.6Hz, 2H), 6.91 (d, J = 8.7Hz, 2H), 5 .72-5.51 (m, 1H), 5.05 (s, 2H), 4.45 (td, J=7.8, 5.2Hz, 1H), 3.94 (q, J=6.7Hz, 1H), 3.03 (dd, J=13.8, 5.1Hz, 1H), 2.94 (dd, J=13.8, 7.7Hz, 1H), 1.12 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ174.23, 172.79, 157.11, 137.19, 130.25, 129.37, 1 28.82, 127.82, 127.73, 114.51, 69.15, 66.97, 52.85, 35.78, 20.97.ESI-MS m / z:[MH] - calcd.for C 19 H 20 NO5 - , 342.1347; Found, 342.1345.

[0107] (R)-3-(4-(benzyloxy)phenyl)-2-(cyclopropaneformamido)propionic acid (18)

[0108]

[0109] Synthetic method is the same as compound 16. Yield: 61%, white powder. 1H NMR (400MHz, DMSO-d6) δ12.62 (s, 1H), 8.36 (d, J = 6.7Hz, 1H), 7.53-7.41 (m, 2H), 7.41-7.35 (m, 2H), 7.35-7.27 (m, 1H), 7.15 (d, J = 6.9Hz, 2 H), 6.92 (d, J=6.8Hz, 2H), 5.06 (s, 2H), 4.51-4.24 (m, 1H), 2.97 (d, J=13.0Hz, 1H), 2.82 (d, J=9.6Hz, 1H), 1.74-1.51 (m, 1H), 0.61 (s, 4H). 13 C NMR (101MHz, DMSO-d6) δ173.33, 172.73, 157.04, 137.22, 130.14, 129.81, 128.44, 127.81, 127.71, 114.49, 69.14, 53.90, 36.07, 13.35, 6.49.ESI-MS m / z:[MH] - calcd.forC 20 H 20 NO4 - , 338.1398; Found, 338.1393.

[0110] (R)-3-(4-((4-fluorobenzyl)oxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (19)

[0111]

[0112] Synthetic method is the same as compound 16. Yield: 53%, white powder. 1 H NMR (400MHz, DMSO-d6). δ12.78 (s, 1H), 7.64 (d, J = 8.2Hz, 1H), 7.50 (d, J = 5.7Hz, 1H) , 7.47 (d, J=5.6Hz, 1H), 7.21 (t, J=8.9Hz, 2H), 7.11 (d, J=8.6Hz, 2H), 6.91 (d, J=8.6 Hz, 2H), 5.50 (s, 1H), 5.03 (s, 2H), 4.43 (td, J=8.0, 5.1Hz, 1H), 3.95 (d, J=5.5Hz, 1H ), 3.01 (dd, J=13.8, 5.1Hz, 1H), 2.94 (dd, J=13.9, 8.0Hz, 1H), 1.15 (d, J=6.8Hz, 3H). 13C NMR (101MHz, DMSO-d6) δ174.66, 173.23, 162.21 (d, J=244.42Hz), 157.44, 133.84 (d, J=3.03Hz), 130.69, 130.41 (d, J=9.09Hz), 129.89, 115.68 (d, J=21.21Hz), 114.96, 68.87, 67.41, 53.28, 36.22, 21.41.ESI-MS m / z:[MH] - calcd.for C 19 H 19 FNO5 - , 360.1253; Found, 360.1250.

[0113] (R)-3-(4-(cyclopropylmethoxy)phenyl)-2-((S)-2-hydroxypropamido)propionic acid (20)

[0114]

[0115] The preparation method is the same as for compound 16. Yield: 56%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.78 (s, 1H), 7.62 (d, J = 8.1Hz, 1H), 7.07 (d, J = 8.5Hz, 2H) , 6.81 (d, J = 8.5Hz, 2H), 5.50 (s, 1H), 4.42 (dt, J = 7.9, 5.1Hz, 1H), 3.94 (d, J = 6.5Hz, 1H), 3.75 (d, J=7.0Hz, 2H), 3.00 (dd, J=13.8, 5.2Hz, 1H), 2.92 (dd, J=13.8, 7.9Hz, 1 H), 1.21-1.16 (m, 1H), 1.15 (d, J=6.7Hz, 3H), 0.61-0.47 (m, 2H), 0.35-0.25 (m, 2H). 13 C NMR (101MHz, DMSO-d6) δ174.20, 172.80, 157.37, 130.19, 128.91, 114.17, 71.89, 66.97, 52.86, 35.78, 20.97, 10.22, 3.11.ESI-MS m / z: [MH] - calcd.for C 16 H 20 NO5 - , 306.1347; Found, 306.1340.

[0116] (R)-2-((S)-2-hydroxypropamido)-3-(4-methoxyphenyl)propionic acid (21)

[0117]

[0118] The preparation method is the same as for compound 16. Yield: 58%, white powder. 1 H NMR (400MHz, DMSO-d6) δ12.78 (s, 1H), 7.63 (d, J = 7.6Hz, 1H), 7.10 (d, J = 7.7Hz, 2H), 6.83 (d, J = 7.6Hz, 2H), 5.50 (s, 1H), 4.53-4.33 (m, 1H), 4.00-3.90 (m, 1H), 3.71 (s, 3H), 2.99 (d, J=7.7Hz, 1H), 2.95 (d, J=7.7Hz, 1H), 1.15 (d, J=6.4Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ174.22, 172.80, 157.98, 130.23, 129.09, 113.65, 66.97, 54.97, 52.88, 35.77, 20.97.ESI-MS m / z: [MH] - calcd.forC 13 H 16 NO5 - ,266.1034;Found,266.1030.0

[0119] Example 2: Study on in vivo appetite-suppressing activity

[0120] Five-week-old male C57BL / 6 mice, weighing 16-20g, were purchased from the Guangdong Provincial Laboratory Animal Center (License No.: SCXK(Y)2022-0002) and housed in an SPF-protected environment. Throughout the experiment, the temperature and relative humidity in the animal room were controlled at 23±2℃ and 50±10%, respectively, with a constant 12-hour light / dark cycle. All mice underwent a one-week acclimatization period before the start of the experiments. Unless otherwise specified, mice were housed under specific pathogen-free conditions and had free access to food and water. The Ethics Committee of Guangdong Pharmaceutical University approved all animal-related procedures and followed the Chinese Regulations on the Management of Laboratory Animals and the Guidelines for the Care and Use of Laboratory Animals issued by the National Institutes of Health (NIH Publication No. 85-23, revised in 2011).

[0121] Mice were randomly divided into a control group and a test compound group, with six mice in each group. Both the control and test compound groups were fed a high-fat diet (Dyets, D12492, containing 60% fat) and normal water intake. After acclimatization, the mice were administered the compound intraperitoneally daily. The control group received water for injection, while the test compound group received 50 mg / kg of the compound. Administration continued for 7 days, and the cumulative appetite suppression rate was calculated. The results of the in vivo appetite-suppressing activity are shown in Table 1. The results indicate that the compound of this invention has good appetite-suppressing activity, suggesting that it has weight-loss potential.

[0122] Table 1: In vivo appetite-suppressing activity

[0123]

[0124] Example 3: In vivo oral administration study on anti-obesity activity

[0125] C57BL / 6 mice were randomly divided into four groups: normal group, model group, compound 1 (150 mg / kg) group, compound 18 (50 mg / kg) group, compound 18 (150 mg / kg) group, and compound 18 (300 mg / kg) group, with six mice in each group. The normal group was given a normal basal diet, while the other groups were fed a high-fat diet (Dyets, D12492, containing 60% fat) and normal drinking water. Starting from week 17, the mice were administered the following medications daily by gavage: 0.5% sodium carboxymethyl cellulose (CMC-Na), compound 1 (150 mg / kg), compound 18 (50 mg / kg), compound 18 (150 mg / kg), and compound 18 (300 mg / kg) for 4 weeks. Mice were fasted for 12 hours prior to sampling (with free access to water). They were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (50 mg / kg). Blood was then collected via the fundus venous plexus. Mice were euthanized by cervical dislocation and fixed on a rat platform. The skin, subcutaneous tissue, and peritoneum were sequentially incised along the midline of the abdomen to expose the abdominal cavity. Liver tissue was removed, washed with physiological saline, and then a suitable size of liver tissue from the left lobe was uniformly cut and fixed in a tissue fixation solution. (A 1×1×0.5 cm sample was taken from the right lobe of the liver, 5 mm from the edge.) 3 Large and small liver tissues were fixed in 4% paraformaldehyde for paraffin embedding and HE staining. The remaining tissues were placed in cryovials, sealed in liquid nitrogen, and stored at -80°C for later use. Experimental results are attached. Figure 1 and 2 .

[0126] Experimental results showed that compounds 1 and 18 significantly reduced body weight, liver weight, and white adipose tissue weight in diet-induced obese (DIO) mice. HE pathological sections revealed that this model induced hepatic steatosis and lobular inflammation. Compared with the model group, compound 1 showed reduced lipid vacuoles and no inflammatory cell infiltration. After treatment with compound 18, lipid vacuoles were significantly reduced, hepatocyte morphology was normal, and no inflammatory cell infiltration was observed; its efficacy was superior to the control compound 1 (see attached). Figure 1 ).

[0127] Furthermore, compared with the normal group, the levels of total triglycerides (TG), total cholesterol (TC), and serum TG were significantly elevated in the HFD-induced DIO mouse model. Compound 1 significantly reduced total TC in the liver, while compound 18 significantly reduced total TG, total TC, and serum TG levels in the liver, with compound 18 showing a stronger effect than compound 1. This suggests that oral administration of compound 18 has a good effect on improving obesity, and its oral efficacy is superior to that of the control compound 1 (see appendix). Figure 2 ).

[0128] Example 4: In vivo oral administration study of anti-nonalcoholic steatohepatitis (NASH) activity

[0129] C57BL / 6 mice were randomly divided into four groups: normal control group, model group, obeticholic acid (OCA) treatment group, compound 18 (50 mg / kg) group, compound 18 (150 mg / kg) group, and compound 18 (300 mg / kg) group, with six mice in each group. The normal control group was given a normal basal diet and drinking water, while the other groups were fed a Western diet (Dyets, D18061501, containing 21.1% fat, 41% fructose, and 1.25% cholesterol) and a sugar solution (23.1 g / L fructose: 18.9 g / L glucose). In addition, except for the normal control group, the other groups were intraperitoneally injected with 0.3 μL / g CCl4 oil solution once a week for 12 weeks to induce the NASH model. Starting from week 9, mice were administered medication daily via gavage. The normal control group and the model group received 0.5% sodium carboxymethyl cellulose (CMC-Na), the OCA group (20 mg / kg), the compound 18 group (50 mg / kg), the compound 18 group (150 mg / kg), and the compound 18 group (300 mg / kg) for 4 weeks. Half an hour after the last administration, a 0.3 μL / g CCl4 oil solution was injected. Mice were fasted for 12 hours before sampling (free access to water), and were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (50 mg / kg). Blood was then collected via the fundus venous plexus. Mice were euthanized by cervical dislocation, fixed on a rat table, and the skin, subcutaneous tissue, and peritoneum were sequentially cut along the midline of the abdomen to expose the abdominal cavity. Liver tissue was removed, washed with physiological saline, and then a suitable size of liver tissue from the left lobe was uniformly cut and fixed in a tissue fixation solution. (A 1×1×0.5 cm sample was taken from the right lobe of the liver, 5 mm from the edge).3 Large and small liver tissues were fixed in 4% paraformaldehyde for paraffin embedding and HE staining. The remaining tissues were placed in cryovials, sealed in liquid nitrogen, and stored at -80°C for later use. Experimental results are attached. Figure 3 and 4 .

[0130] Experimental results showed that HE pathological sections induced typical histological features of NASH, including steatosis, lobular inflammation, and hepatocyte ballooning degeneration. Compared with the model group, the OCA treatment group showed a significant reduction in lipid vacuoles and no dense inflammatory cell infiltration. After treatment with compound 18, inflammation was significantly reduced, hepatocyte morphology was normal, there was no ballooning degeneration, and the cells were arranged more neatly, with only some slight lipid vacuoles observed. NAS score results showed that the therapeutic effect of compound 18 was comparable to that of OCA (see appendix). Figure 3 ).

[0131] Furthermore, compared with the normal group, the levels of total triglycerides (TG), total cholesterol (TC), and serum TG in the NASH mouse model were significantly elevated, while these indicators were significantly reduced after treatment with OCA and compound 18, with comparable effects. Because the fibrosis promoter CCl4 causes some damage to the mouse liver, the serum AST and ALT levels in the model group were significantly elevated compared to the normal group. In contrast, AST and ALT levels in the treatment group were significantly reduced, suggesting that OCA and compound 18 have a good protective effect on liver function and can be used to prevent and treat liver fibrosis. (Appendix) Figure 4 This indicates that the oral administration of compound 18 has an anti-NASH efficacy comparable to that of the marketed drug OCA, and has broad application prospects.

[0132] Example 5: Study on antidiabetic activity of oral administration in vivo

[0133] C57BL / 6 mice were fed a normal basal diet and drinking water, and after one week of acclimatization, diabetes analysis was performed. STZ was administered intraperitoneally at a dose of 50 mg / kg for three consecutive days. Blood glucose levels were measured after each injection. Mice with fasting blood glucose greater than 11.1 mmol / L and random blood glucose greater than 16.7 mmol / L were randomly divided into four groups: normal control group, model group, empagliflozin group, and compound 18 (150 mg / kg) group, with six mice in each group. After grouping, mice were administered 0.5% carboxymethyl cellulose sodium (CMC-Na), empagliflozin (20 mg / kg), or compound 18 (150 mg / kg) by gavage for two weeks, respectively. The results of blood glucose improvement are shown in Table 2. The results indicate that compound 18 of this invention has a good blood glucose improvement effect, and its efficacy is even superior to that of the marketed drug empagliflozin.

[0134] Table 2: Blood glucose levels in mice

[0135]

[0136] Example 6: Study on tolerance to hypoxia in normobaric hypoxic environment

[0137] Five-week-old male Kunming mice weighing 18-22g were randomly divided into four groups: a model group, a test compound 1 group, a test compound 8 group, and a test compound 14 group, with six mice in each group. The model group was administered 0.5% sodium carboxymethyl cellulose (CMC-Na) by gavage. The other test compounds were administered compound 1 (150 mg / kg), compound 8 (150 mg / kg), and compound 14 (150 mg / kg) by gavage, respectively, for three consecutive days. After this, each mouse was placed individually in a 125ml wide-mouth bottle, sealed with a rubber stopper, and the asphyxiation time was closely observed and recorded. The results are shown in Table 3. After the experiment, the mice were anesthetized and euthanized. Brain tissue was immediately dissected, placed in cryovials, sealed in liquid nitrogen, and then stored at -80℃. Subsequent assay kits were used to detect the levels of MDA, SOD, and GSH-Px in the brain tissue. The experimental results are shown in Table 4.

[0138] Table 3: Duration of Asphyxiation in Animals

[0139]

[0140] Table 4: Content of MDA, SOD, and GSH-Px in brain tissue

[0141]

[0142] Experimental results showed that compound 14 significantly increased the asphyxiation time in mice under closed hypoxic conditions, while compounds 1, 8, and 14 significantly increased the levels of SOD and GSH-Px. Compounds 1 and 14 significantly decreased the MDA content in brain tissue. This indicates that compound 14 can enhance the hypoxia tolerance of mice under normobaric hypoxia, suggesting its potential for treatment of cerebral ischemia, myocardial infarction, and altitude sickness.

[0143] Example 7: Study on tolerance to hypoxia in low-pressure hypoxic environments (simulating low-pressure, hypoxic environments at high altitudes)

[0144] Kunming mice were randomly divided into four groups: a model group, a test compound 1 group, a test compound 10 group, and a test compound 14 group, with six mice in each group. The model group was administered 0.5% sodium carboxymethyl cellulose (CMC-Na) by gavage. The other test compounds were administered by gavage to compound 1 (150 mg / kg), compound 10 (150 mg / kg), and compound 14 (150 mg / kg), respectively, for three consecutive days. After this, each mouse was individually placed in a 125 ml wide-mouth bottle containing approximately 5 g of soda lime, sealed with a rubber stopper, and the asphyxiation time was closely observed and recorded. The results are shown in Table 5. After the experiment, the mice were anesthetized and euthanized. Brain tissue was immediately dissected and stored in cryovials under liquid nitrogen at -80°C. Subsequent assay kits were used to detect the levels of MDA, SOD, and GSH-Px in the brain tissue. The experimental results are shown in Table 6.

[0145] Table 5: Duration of Asphyxiation in Animals

[0146]

[0147] Table 6: Content of MDA, SOD, and GSH-Px in Brain Tissue

[0148]

[0149] Experimental results showed that compound 14 significantly increased the survival time of mice in a closed, low-pressure, hypoxic environment, while compounds 1 and 14 significantly increased the levels of SOD and GSH-Px and significantly decreased the level of MDA in brain tissue. This indicates that compounds 1 and 14 can enhance the hypoxia tolerance of mice in low-pressure, hypoxic environments and have the potential to prevent and treat altitude sickness.

[0150] Example 8: In vivo study on anti-hyperuricemia activity

[0151] Kunming mice were randomly divided into four groups: a model group, a test compound 1 group, a test compound 8 group, a test compound 18 group, and a test compound 19 group, with six mice in each group. At -1 hour, the animals were administered 0.5% sodium carboxymethyl cellulose (CMC-Na), compound 1 (150 mg / kg), compound 8 (150 mg / kg), compound 18 (150 mg / kg), and compound 19 (150 mg / kg) by gavage, respectively. At 0 min, hypoxanthine 200 mg / kg was injected intraperitoneally, and potassium oxonate 120 mg / kg was injected subcutaneously to induce the model. Blood uric acid levels were measured by tail sampling at 1, 2, and 4 hours post-modeling. The results are shown in Table 7.

[0152] Table 7: Blood uric acid levels in mice

[0153]

[0154] Experimental results show that compounds 1, 8, and 19 have good anti-hyperuricemia activity.

[0155] Example 9: In vivo study on anti-acute alcohol poisoning activity

[0156] Kunming mice were randomly divided into four groups: a model group, a test compound 1 group, a test compound 8 group, a test compound 18 group, and a test compound 19 group, with 10 mice in each group. At -1 hour, the animals were administered 0.5% sodium carboxymethyl cellulose (CMC-Na), compound 1 (150 mg / kg), compound 8 (150 mg / kg), compound 18 (150 mg / kg), and compound 9 (150 mg / kg) by gavage, respectively. At 0 min, each mouse was administered 0.025 ml / g of 52% baijiu (a type of Chinese liquor) by gavage. The disappearance of the righting reflex was used as the indicator of intoxication. The number of intoxicated and dead mice was recorded, and the intoxication rate and mortality rate were calculated. The results are shown in Table 8.

[0157] Table 8: Intoxication rate and mortality in mice

[0158]

[0159] Experimental results show that compounds 1, 18, and 19 have good anti-acute alcohol poisoning activity.

[0160] Example 10: In vivo study of anti-acetaminophen-induced liver injury activity

[0161] Kunming mice were randomly divided into three groups: a normal group, a model group, a test compound 1 group, a test compound 18 group, and a test compound 19 group, with six mice in each group. The normal group and the model group were administered 0.5% sodium carboxymethyl cellulose (CMC-Na) by gavage, while the other test compound groups were administered compound 1 (150 mg / kg), compound 18 (150 mg / kg), and compound 19 (150 mg / kg) by gavage, respectively, for three consecutive days. On the evening of the third day, the mice were fasted for 12 hours but allowed normal access to water. After fasting, except for the normal group, each mouse was intraperitoneally injected with 300 mg / kg of APAP solution. Ten hours later, they were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (50 mg / kg). Blood was then collected via orbital sampling. Mice were euthanized by cervical dislocation and fixed on a mouse table. The skin, subcutaneous tissue, and peritoneum were sequentially cut along the midline of the abdomen to expose the abdominal cavity. Liver tissue was removed, washed with physiological saline, and then a suitable size of liver tissue from the left lobe was uniformly cut and fixed in tissue fixation solution. (A 1×1×0.5 cm section was taken from the right lobe of the liver, 5 mm from the edge.) 3 Large and small liver tissues were fixed in 4% paraformaldehyde for paraffin embedding and HE staining. The remaining tissues were placed in cryovials, sealed in liquid nitrogen, and stored at -80°C for later use. Experimental results are attached. Figure 5 .

[0162] Experimental results showed that HE pathological sections indicated that this model induced liver damage in mice. Treatment with compounds 1, 18, and 19 significantly improved liver damage, and compared with the model group, serum AST and ALT levels in mice were significantly decreased, indicating that compounds 1, 18, and 19 had good hepatoprotective effects. (See appendix) Figure 5 ).

Claims

1. A compound of the general formula (I): ###0001### (I) wherein: R1 is selected from the group consisting of: ###0002### R2 is selected from the group consisting of: ###0003### 2. The compound of general formula (I) as defined in claim 1 is selected from the group consisting of: (S)-3-(4-(benzyloxy)phenyl)-2-propionamidopropanoic acid; (S)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropanamidopropionic acid; (R)-3-(4-(benzyloxy)phenyl)-2-((R)-2-hydroxy-3-methylbutanamidopropionic acid; (R)-3-(4-(benzyloxy)phenyl)-2-propionamidopropanoic acid; (R)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropanamidopropionic acid; (R)-3-(4-(benzyloxy)phenyl)-2-(cyclopropanecarboxamidopropionic acid; (R)-3-(4-((4-fluorobenzyl)oxy)phenyl)-2-((S)-2-hydroxypropanamidopropionic acid. wherein 3. A pharmaceutical composition comprising a compound as defined in any one of claims 1-2 and a suitable carrier or excipient. wherein R is optionally selected from hydrogen, halogen; 4. Use of the compound (S)-3-(4-(benzyloxy)phenyl)-2-((S)-2-hydroxypropanamidopropionic acid as defined in claim 2 for the manufacture of a medicament for the prevention or / and treatment of at least one of the following diseases: high altitude sickness, brain hypoxia, brain ischemia, myocardial infarction.

5. Use of a compound as defined in any one of claims 1-2, or a pharmaceutical composition as defined in claim 3, for the manufacture of a medicament for the prevention or / and treatment of at least one of the following diseases: obesity, non-alcoholic fatty liver, alcoholism, alcoholic steatohepatitis, drug-induced liver injury, dyslipidemia, liver cirrhosis, liver failure, diabetes, diabetic complications, hyperuricemia. ​ ​ ​ ​ ​ ​ ​ ​ ​ ​

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