An indol ethylamine derivative, its preparation method and application

Abstract

The application discloses an indole ethylamine derivative, a preparation method and application thereof, and belongs to the technical field of organic synthesis. The structural formula of the indole ethylamine derivative is shown in formula (I) or formula (II). The indole ethylamine derivative is prepared from tryptamine or 6-halogen-substituted indole, and has the advantages of simple preparation method, high yield, easy-to-obtain raw materials and large-scale production. The indole ethylamine derivative can be used as a double-effect regulator of peroxisome proliferator-activated receptor PPAR alpha and carnitine palmitoyltransferase CPT1, has good effects of resisting fat accumulation, resisting oxidation and resisting inflammation, and has certain potential in preparation of a medicine for treating non-alcoholic fatty liver disease.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of organic synthesis, in particular to an indole ethylamine derivative and a preparation method and application thereof. BACKGROUND

[0002] NAFLD (nonalcoholic fatty liver disease) refers to acquired metabolic stress liver injury with excessive deposition of fat in liver cells as the main feature, except for alcohol and other clear liver damage factors. In recent years, with the development of social economy and the change of lifestyle, NAFLD has become the most common chronic liver disease in the world, affecting people's health, and has become an important public health problem in China in recent years. NAFLD is harmful and there is no effective targeted drug in clinical practice. Finding effective drugs for treating NAFLD is an urgent need for clinical medicine.

[0003] Excessive accumulation of triglycerides, free fatty acids, diglycerides, oxidized sterols and other lipids in hepatocytes is the main factor of NAFLD. The essence of NAFLD is the imbalance of lipid metabolism in the liver, mainly manifested in the increase of lipid entering the liver and the decrease of metabolized lipid. Peroxisome proliferator-activated receptor alpha (PPARα) is an important factor in regulating metabolic syndrome, and also an important factor in regulating lipid metabolism. PPARα is highly expressed in the liver and is involved in fatty acid uptake, binding, oxidation and lipid transport. PPARα can directly regulate carnitine palmitoyltransferase 1 (CPT1). CPT1 is a rate-limiting enzyme for fatty acid beta-oxidation, thereby regulating the process of hepatic fatty acid beta-oxidation, which provides a new idea for the treatment of NAFLD. Therefore, we speculate that the dual activator of PPARα / CPT1 has great practical significance in the treatment of NAFLD.

[0004] Indole derivatives exist widely in nature, most of which have biological activity and are widely used in the fields of pesticides, medicines, dyes, feed, food and additives. Obtaining indole derivatives by organic synthesis method is also a current research hotspot. Chinese patent document with publication number CN113292476A discloses sulfanilamide indole derivatives, a preparation method and application thereof. The general formula of the sulfanilamide indole derivative is shown in the following formula (left). Such compounds have good inhibitory activity against gram-positive bacteria and gram-negative bacteria, especially against Staphylococcus aureus, and are less likely to develop drug resistance and have low cytotoxicity. Chinese patent document with publication number CN115368289A discloses an antitumor bisindole methane derivative compound, a preparation method and application thereof. The structural formula of the bisindole methane derivative is shown in the following formula (right). Such bisindole methane has significant antitumor activity and can efficiently kill multiple types of cancer cells.

[0005]

[0006] Therefore, in view of the current situation of the treatment of NAFLD, it is a research direction worth exploring to develop new chemical drugs with high efficiency and low toxicity. SUMMARY

[0007] The present application provides an indole ethylamine derivative, which has a simple preparation method, raw materials are easy to obtain, can be used as a dual-effect regulator of peroxisome proliferator-activated receptor PPARα and carnitine palmitoyltransferase CPT1, and has good effects of anti-fat accumulation, anti-oxidation and anti-inflammation.

[0008] The specific technical solutions are as follows:

[0009] An indole ethylamine derivative, the structural formula of which is shown in formula (I) or formula (II):

[0010]

[0011] In formula (I), R1 is hydroxyl or alkoxy-substituted amino; in formula (II), X is halogen; R2 is any one of the following, is a substituted position;

[0012]

[0013] The present application also provides a preparation method of the indole ethylamine derivative, specifically, when the indole ethylamine derivative has the structural formula shown in formula (I), the preparation method comprises the following steps:

[0014] S01 adding triethylamine and di-tert-butyl dicarbonate in sequence in a solution of tryptamine to obtain compound 1 by reaction;

[0015] S02 reacting compound 1 with an oxidant 2,3-dichloro-5,6-dicyano-p-benzoquinone to obtain compound 2;

[0016] S03 adding sodium hydride to a solution of compound 2, and then adding p-methoxybenzyl chloride to obtain compound 3 by reaction;

[0017] S04 reacting compound 3 and hydrogenated diisobutylaluminum in an organic solvent to obtain an indole ethylamine derivative with R1 being hydroxyl;

[0018] and / or, adding alkoxyamine hydrochloride and triethylamine in sequence to an organic solution of the indole ethylamine derivative with R1 being hydroxyl to obtain an indole ethylamine derivative with R1 being alkoxy-substituted amino.

[0019] Preferably, in step S01, the reaction temperature is 10-50℃, and the reaction time is 1-5h.

[0020] Preferably, in step S02, tetrahydrofuran and water are used as the solvent of the reaction system, the reaction temperature is -10-60℃, and the reaction time is 1-6h.

[0021] Preferably, in step S03, the reaction temperature is -10-50℃, the reaction time is 0.5-5h, and the p-methoxybenzyl halide is preferably 4-methoxybenzyl bromide.

[0022] Preferably, in step S04, the reaction temperature of compound 3 and diisobutylaluminum hydride is -100-20℃, and the reaction time is 0.5-3h; and / or, the reaction temperature of the alkoxyamine hydrochloride, the indoleethylamine derivative with hydroxyl-substituted R1, and triethylamine is 80-150℃, and the reaction time is 15-30h.

[0023] The present application also provides a preparation method of the indoleethylamine derivative, specifically, when the indoleethylamine derivative has the structural formula shown in formula (II), the preparation method comprises:

[0024] S11 mixing sodium hydride with a solution of 6-halogen-substituted indole, stirring, and then adding p-methoxybenzyl halide to obtain compound 6;

[0025] S12 reacting compound 6 with methylmalonyl chloride under the action of aluminum chloride to obtain compound 7, and adding p-acetamidobenzenesulfonyl azide and triethylamine to a solution of compound 7 to obtain the indoleethylamine derivative with R2 as shown in formula (III);

[0026] and / or, reacting the indoleethylamine derivative with R2 as shown in formula (III) and the indoleethylamine derivative with R1 being alkoxy-substituted amino in trifluorotoluene to obtain intermediate product 1, adding 2,6-dimethylaniline and trimethylsilyl trifluoromethanesulfonate to a dichloromethane solution of intermediate product 1 to obtain the indoleethylamine derivative with R2 as shown in formula (IV).

[0027] Preferably, in step S11, the 6-halogen-substituted indole is 6-bromindole, the p-methoxybenzyl halide is 4-methoxybenzyl bromide, the reaction temperature is -20-60℃, and the reaction time is 3-9h.

[0028] Preferably, in step S12, the reaction temperature of compound 6 and methyl malonyl chloride is -20-50℃, the reaction time is 8-15h; the reaction temperature of compound 7 and p-acetamidobenzenesulfonyl azide, triethylamine is -20-60℃, the reaction time is 6-18h; and / or, the reaction temperature of indole ethylamine derivative of R2 as shown in formula (III), indole ethylamine derivative of R1 being alkoxy-substituted amino in trifluorotoluene is 50-150℃, the reaction time is 2-8h, the reaction temperature of intermediate product 1 and 2,6-dimethylaniline, trimethylsilyl trifluoromethanesulfonate is -20-40℃, the reaction time is 1-5h.

[0029] The present application also provides a medicine comprising the indole ethylamine derivative, and the medicine is used for resisting fat accumulation, resisting oxidation and / or resisting inflammation.

[0030] Experiments prove that the indole ethylamine derivative can be used as a double-effect regulator of peroxisome proliferator-activated receptor PPARα and carnitine palmitoyl transferase CPT1, and has good effects of resisting fat accumulation, resisting oxidation and resisting inflammation.

[0031] Compared with the prior art, the present application has the beneficial effects that:

[0032] (1) The present application designs a series of target structures according to the principle of optimizing the mother nucleus structure, and synthesizes four kinds of indole ethylamine derivatives by using convenient and easily obtained raw materials, the structure and spectrum data (including single crystal data) and activity research of which have not been reported, and the preparation method of the indole ethylamine derivative provided by the present application is simple, the yield is high, the raw materials are easy to obtain, and the indole ethylamine derivative can be produced on a large scale.

[0033] (2) The indole ethylamine derivative can be used as a double-effect regulator of peroxisome proliferator-activated receptor PPARα and carnitine palmitoyl transferase CPT1, and has good effects of resisting fat accumulation, resisting oxidation and resisting inflammation, and has certain potential in preparing a medicine for treating non-alcoholic fatty liver disease. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 It is a preparation route map of the indole ethylamine derivative.

[0035] Figure 2 It is an influence of the indole ethylamine derivative on AML12 cells induced by oleic acid, wherein A is an influence of the four kinds of indole ethylamine derivatives on the survival rate of AML12 cells, B is an influence of the four kinds of indole ethylamine derivatives on the triglyceride content of AML12 cells induced by oleic acid, C is an influence of compound 9 at different concentrations on the survival rate of AML12 cells, D is an influence of compound 9 at different concentrations on the triglyceride content of AML12 cells induced by oleic acid, *** indicates a statistical difference of P<0.001, and ns indicates no statistical difference.

[0036] Figure 3 Effect of compound 9 on oil red O staining of AML12 cells induced by oleic acid.

[0037] Figure 4 Effect of compound 9 on Bodipy fluorescence staining of AML12 cells induced by oleic acid.

[0038] Figure 5 Effect of compound 9 on mRNA expression levels of PPARa / CPT1 pathway and lipid metabolism related genes in AML12 cells induced by oleic acid. *, **, *** represent statistical differences of P<0.05, P<0.01, P<0.001, respectively.

[0039] Figure 6 Effect of compound 9 on protein expression levels of PPARa / CPT1 pathway and lipid metabolism related genes in AML12 cells induced by oleic acid. DETAILED DESCRIPTION

[0040] The present application is further illustrated by the following examples in conjunction with the accompanying drawings and examples. It should be understood that these examples are only used to illustrate the present application, and are not used to limit the scope of the present application.

[0041] In the examples, the preparation route of the indolylethylamine derivative is as shown in Figure 1

[0042] Example 1

[0043]

[0044] (1) To a solution of tryptamine (5.00 g, 31.23 mmol) in tetrahydrofuran (THF, 15 mL) was added triethylamine (NEt3, 6.5 mL, 46.84 mmol), followed by a solution of di-tert-butyl dicarbonate ((Boc)2O, 8.60 mL, 37.48 mmol) in THF (20 mL) and stirred at 25 °C for 1 h. After the reaction was completed, the mixture was treated by adding water (50 mL) and ethyl acetate (30 mL). The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 30 mL). The combined extracts were washed with brine (60 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluted with n-hexane / ethyl acetate (7:3) to give compound 1 (7.96 g, yield 98%).

[0045] The characterization data of compound 1 are as follows:

[0046] 1 ​H NMR (400 MHz, Chloroform-d) δ 8.54 (s, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.22 (ddd, J = 8.2, 6.9, 1.2 Hz, 1H), 7.14 (ddd, J = 8.1, 7.0, 1.2 Hz, 1H), 6.99 (s, 1H), 4.75 (s, 1H), 3.49 (t, J = 6.7 Hz, 2H), 2.97 (t, J = 6.7 Hz, 2H), 1.48 (s, 9H).

[0047]

[0048] (2) Compound 1 (4.0 g, 15.3 mmol) was dissolved in tetrahydrofuran / water (16:1, 34 mL) and 2,3-dichloro-5,6-dicyanoquinone (DDQ, 6.91 g, 30.6 mmol, 2.0 eq) was added to it at 0 °C and the reaction was further stirred at 25 °C for 2 h. After completion of the reaction, water (50 mL) and ethyl acetate (30 mL) were added to the mixture. The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 30 mL). The combined extracts were washed successively with saturated aqueous NaHC03(4 x 25 mL) and brine (60 mL), dried over anhydrous Na2S04and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluting with n-hexane / ethyl acetate (2:1) to obtain compound 2 (2.99 g, yield 71 %).

[0049] Characterization data of compound 2 are as follows:

[0050] 1 H NMR (400 MHz, DMSO-d6) δ 12.01 (1H, br s, NH), 8.42 (d, J = 2.8, 1H), 8.22 (d, J = 6.1, 1H), 7.51 (d, J = 7.3 Hz, 1H), 7.41 - 7.13 (m, 2H), 7.02 (t, J = 7.0 Hz, 1H), 4.32 (d, J = 6.0 Hz, 2H), 1.43 (s, 9H).

[0051] 13 C NMR (100 MHz, DMSO-d6) δ 190.89, 156.08, 136.50, 133.36, 125.50, 122.90, 121.85, 121.30, 114.07, 112.22, 77.95, 46.95, 28.30.

[0052]

[0053] (3) To a solution of compound 2 (1.0 g, 3.60 mmol) in THF (15 mL) was added sodium hydride (60% dispersion in mineral oil, 4.3 mmol) in portions at 0 °C. After 30 min, 4-methoxybenzyl bromide (4.0 mmol) was added to the reaction mixture. The resulting mixture was further stirred at 25 °C for 1 h. After completion of the reaction, water (20 mL) and ethyl acetate (20 mL) were added to the mixture. The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 20 mL). The combined extracts were washed with brine (60 mL), dried over anhydrous Na2S04and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluted with n-hexane / ethyl acetate (4:1) to afford compound 3 (0.92 g, yield 64%).

[0054] Characterization data of compound 3 are as follows:

[0055] 1 H NMR (400 MHz, Chloroform-d) δ 8.33 (dd, J = 7.4, 1.8 Hz, 1H), 7.74 (d, J = 2.6 Hz, 1H), 7.38 - 7.34 (m, 1H), 7.29 (dt, J = 4.4, 2.4 Hz, 2H), 7.12 (dd, J = 8.8, 3.0 Hz, 2H), 6.89 - 6.85 (m, 2H), 5.69 (s, 1H), 5.20 (s, 2H), 4.48 (s, 2H), 3.79 (s, 3H), 1.49 (s, 9H).

[0056] 13 C NMR (100 MHz, Chloroform-d) δ 189.21, 159.79, 156.06, 137.05, 134.16, 129.07, 127.09, 126.43, 123.77, 123.10, 122.46, 114.65, 114.33, 110.46, 79.71, 55.42, 50.50, 47.64, 28.50.

[0057] HR-ESI-MS (m / z): 395.1935 (calcd m / z 395.1965 [M+H] + ) Molecular Formula C 23 H 26 N2O4.

[0058]

[0059] (4) To a solution of compound 3 (500 mg, 1.3 mmol) in THF (10 mL) was added a solution of diisobutylaluminum hydride (DIBAL-H, 2.6 mmol) in n-hexane (2.6 mL) under argon at -78 °C and the reaction stirred for 1 h. After completion of the reaction, saturated aqueous Na2KCO3solution (5 mL) was added and the reaction mixture stirred vigorously for 30 min. The mixture was treated with water (20 mL) and ethyl acetate (20 mL). The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 20 mL). The combined extracts were washed with brine (40 mL), dried over anhydrous Na2SO4and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluting with n-hexane / ethyl acetate (4:1) to afford compound 4 (0.27 g, yield 55%).

[0060] Characterization data of compound 4 are as follows:

[0061] 1 H NMR (400 MHz, Chloroform-d) δ 7.61 (d, J = 7.9 Hz, 1H), 7.17 (t, J = 7.7 Hz, 1H), 7.08 (t, J = 7.7 Hz, 1H), 7.02 (d, J = 7.9 Hz, 1H), 6.98 (s, 1H), 6.96 (d, J = 8.6 Hz, 2H), 6.71 (d, J = 8.6 Hz, 2H), 5.08 (s, 1H, OH), 5.05 (s, 2H), 5.00 (dd, J = 8.7, 3.6 Hz, 1H), 3.65 (s, 3H), 3.56 - 3.44 (m, 1H), 3.46 - 3.26 (m, 1H), 1.35 (s, 9H).

[0062] 13 C NMR (100 MHz, Chloroform-d) δ 159.06, 155.80, 136.74, 129.08, 128.30, 128.01, 125.49, 121.98, 119.46, 114.13, 114.09, 113.99, 109.88, 79.47, 68.13, 55.18, 49.48, 28.32.

[0063] HR-ESI-MS (m / z): 397.2160 (calcd m / z 397.2122 [M+H]) + ) Molecular Formula C 23 H 28 N2O4.

[0064] IR (KBr) v max3346, 2977, 2933, 1699, 1640, 1514, 1390, 1249, 1175, 1033, 823, 746 cm -1 .

[0065] UV (MeOH) λ max = 195 nm, 210 nm, 250 nm, 310 nm. Melting point: 125-126 °C.

[0066] Example 2

[0067]

[0068] MeONH2 HCI (158 mg, 1.89 mmol) was added to a solution of compound 4 (150 mg, 0.37 mmol) in toluene (5 mL) at room temperature, followed by the addition of NEt3 (0.15 mL, 1.13 mmol) and the mixture was stirred at 110 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature and treated with the addition of water (5 mL) and ethyl acetate (5 mL). The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 5 mL). The combined extracts were washed with brine (10 mL), dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluted with n-hexane / ethyl acetate (2:1) to give compound 5. (90 mg, 55% yield).

[0069] The characterization data of compound 5 are as follows:

[0070] 1 H NMR (400 MHz, Chloroform-d) δ 7.63 (d, J = 7.9 Hz, 1H), 7.20 (t, J = 7.8 Hz, 1H), 7.11 (t, J = 7.8 Hz, 1H), 7.05 (d, J = 7.9 Hz, 1H), 7.03 (s, 1H), 6.99 (d, J = 8.5 Hz, 2H), 6.74 (d, J = 8.5 Hz, 2H), 5.12 (s, 2H), 4.79 (d, J = 7.7 Hz, 1H, NH), 4.39 (t, J = 6.2 Hz, 1H), 3.69 (s, 3H), 3.64 (q, J = 5.8 Hz, 1H), 3.54 - 3.50 (m, 1H), 3.50 (s, 3H), 1.33 (s, 9H).

[0071] 13C NMR (100 MHz, Chloroform-d) δ 159.01, 155.93, 136.42, 129.11, 128.19 (2C), 126.29, 121.96, 121.70, 119.42, 119.18, 114.03 (2C), 112.05, 109.85, 78.93, 62.41, 57.46, 55.07, 49.42, 43.14, 28.30.

[0072] HR-ESI-MS (m / z): 426.2312 (calcd m / z 426.2387 [M+H] + ) Molecular Formula C 24 H 31 N3O4.

[0073] IR (KBr) v max 2976, 2934, 1710, 1612, 1513, 1466, 1392, 1366, 1248, 1173, 1034, 821, 741 cm -1 .

[0074] UV (MeOH) λ max = 200 nm, 230 nm, 290 nm.

[0075] Example 3

[0076]

[0077] To a solution of compound 6 (0.55 g, 1.89 mmol) in 10 mL of anhydrous THF and sodium hydride (60% dispersion in mineral oil, 6.1 mmol) was added 4- methoxybenzyl chloride (PMBBr, 0.81 mL, 5.6 mmol) after stirring at room temperature for 1 h under nitrogen protection and cooling by ice water bath. The reaction mixture was further stirred at 25 °C for 6 h. After the reaction was completed, the reaction mixture was quenched by dropwise addition of water (15 mL) and ethyl acetate (15 mL) was added to the mixture. The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined extracts were washed with brine (30 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluted with n-hexane / ethyl acetate (8:1) to give compound 7 (0.55 g, yield 34%).

[0078] The characterization data of compound 6 are as follows:

[0079] 1H NMR (400 MHz, Chloroform-d) δ 7.38 (d, J = 8.4 Hz, 1H), 7.35 (dd, J = 1.7, 0.8 Hz, 1H), 7.10 (dd, J = 8.4, 1.7 Hz, 1H), 6.98 (d, J = 8.6 Hz, 1H), 6.95 (d, J = 3.2 Hz, 1H), 6.92 (d, J = 8.8 Hz, 2H), 6.72 (d, J = 8.7 Hz, 2H), 6.39 (dd, J = 3.2, 0.9 Hz, 1H), 5.05 (s, 2H), 3.65 (s, 3H).

[0080]

[0081] Compound 6 (580 mg, 1.8 mmol), AICI3(490 mg, 3.6 mmol) and dichloromethane (CH2CI2, 10 mL) were added to a reaction flask, and methyl malonyl chloride (0.23 mL, 2.2 mmol) was added dropwise to the reaction flask at 0 °C, and the reaction was stirred at room temperature 25 °C for 11 hours. After the reaction was completed, dichloromethane (10 mL) and water (15 mL) were added for treatment. The organic phase was separated, and the aqueous layer was extracted with dichloromethane (2 x 15 mL). The combined extracts were washed with brine (20 mL), dried over anhydrous Na2S04, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel, eluted with n-hexane / ethyl acetate (2: 1), to obtain compound 7 (0.25 g, yield 34%).

[0082] The characterization data of compound 7 are as follows:

[0083] 1 H NMR (400 MHz, Chloroform-d) δ 8.15 (d, J = 8.5 Hz, 1H), 7.64 (s, 1H), 7.39 (d, J = 1.7 Hz, 1H), 7.30 (dd, J = 8.5, 1.7 Hz, 1H), 7.02 (d, J = 8.6 Hz, 2H), 6.80 (d, J = 8.6 Hz, 2H), 5.11 (s, 2H), 3.74 (s, 2H), 3.71 (s, 3H), 3.64 (s, 3H).

[0084] 13 C NMR (100 MHz, Chloroform-d) δ 186.33, 168.44, 159.82, 137.96, 135.97, 128.76 (2C), 126.81, 126.46, 125.47, 124.08, 117.59, 116.33, 114.68 (2C), 113.45, 55.44, 52.58, 50.56, 47.12.

[0085] HR-ESI-MS (m / z): 438.0433 (calcd m / z 438.0311 [M+2+Na] + ) Formula C 20 H 18 BrNO4.

[0086] IR (KBr) v max 2929, 1611, 1513, 1462, 1313, 1249, 1175, 1034, 890, 802, 718 cm -1 .

[0087] UV (MeOH) λ max = 200 nm, 230 nm, 295 nm.

[0088] Example 4

[0089]

[0090] Compound 7 (150 mg, 0.36 mmol) was dissolved in acetonitrile (CH3CN, 4 mL), to which p-acetamidobenzenesulfonyl azide p-ABSA (130 mg, 0.54 mmol) was added and it was cooled to 0 °C, NEt3(0.12 mL, 0.9 mmol) was added, the reaction mixture was brought to 25 °C and stirred for 12 h. After the end of the reaction, the resulting mixture was treated by adding ethyl acetate (4 mL) and water (5 mL). The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 4 mL). The combined extracts were washed with brine (10 mL), dried over anhydrous Na2S04and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluted with n-hexane / ethyl acetate (2:1) to give compound 8 (43 mg, yield 27%).

[0091] The characterization data of compound 8 are as follows:

[0092] 1 H NMR (400 MHz, Chloroform-d) δ 8.33 (s, 1H), 8.23 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 1.7 Hz, 1H), 7.36 (dd, J = 8.5, 1.7 Hz, 1H), 7.09 (d, J = 8.5 Hz, 2H), 6.85 (d, J = 8.5 Hz, 2H), 5.22 (s, 2H), 3.82 (s, 3H), 3.77 (s, 3H).

[0093] 13C NMR (100 MHz, CDC13) δ 177.63, 162.38, 159.62, 137.57, 137.11, 128.46 (2C), 127.32, 126.95, 126.03, 123.98, 116.98, 114.53 (2C), 113.79, 113.35, 55.39, 52.24, 50.62.

[0094] HR-ESI-MS (m / z): 442.0365 (calcd m / z 442.0397 [M+H]) + ) molecular formula C 20 H 16 BrN3O4.

[0095] IR (KBr) v max 3140, 2954, 2838, 2144, 1720, 1611, 1578, 1514, 1468, 1369, 1294, 1249, 1177, 1105, 1034, 937, 866, 812, 743 cm -1 .

[0096] UV (MeOH) λ max = 195 nm, 220 nm, 250 nm, 280 nm, 320 nm.

[0097] Example 5

[0098]

[0099] Compound 5 (40 mg, 0.1 mmol) and compound 8 (49 mg, 0.12 mmol) were added to trifluorotoluene (PhCF3, 2 mL) under nitrogen and heated to 110 °C for 5 h. After the reaction was completed, the mixture was cooled to room temperature and treated by adding ethyl acetate (5 mL) and water (5 mL). The organic phase was separated and the aqueous layer was extracted with ethyl acetate (2 x 5 mL). The combined extracts were washed with brine (10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluted with n-hexane / ethyl acetate (2:1) to give intermediate product 1 (30 mg, 37% yield).

[0100] Intermediate 1 (40 mg, 0.05 mmol) was dissolved in dichloromethane (CH2Cl2, 2 mL) under nitrogen, the resulting solution was cooled to 0 °C and 2,6-lutidine (0.023 mL, 0.2 mmol) and trimethylsilyl trifluoromethanesulfonate (TMSOTf, 0.026 mL, 0.15 mmol) were added successively, the reaction mixture was allowed to warm to room temperature 25 °C and stirred for 3.5 h. After the end of the reaction, dichloromethane (3 mL) and water (3 mL) were added. The organic phase was separated and the aqueous layer was extracted with dichloromethane (2 x 3 mL). The combined extracts were washed successively with saturated aqueous sodium bicarbonate solution (3 mL) and brine (6 mL), dried over anhydrous Na2SO4and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel eluting with n-hexane / ethyl acetate (1 : 1) to give compound 9 (13 mg, 57% yield).

[0101] The characterization data of compound 9 are as follows:

[0102] 1 H NMR (400 MHz, DMSO-d6) δ 7.74 (d, J = 1.8 Hz, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.49 (s, 1H), 7.20 (d, J = 8.7 Hz, 2H), 7.16 (d, J = 8.5, 1.8 Hz, 1H), 6.88 (d, J = 8.7 Hz, 2H), 5.33 (s, 2H), 4.74 (s, 1H), 3.70 (s, 3H), 3.64 (s, 3H), 3.56 (s, 3H).

[0103] 13 C NMR (100 MHz, DMSO-d6) δ 168.98, 164.06, 158.70, 136.52, 136.31, 129.59, 129.17, 128.77 (2C), 126.15, 121.92, 121.31, 114.35, 114.02 (2C), 112.98, 107.20, 63.19, 55.08, 52.38, 48.50, 46.76.

[0104] HR-ESI-MS (m / z): 463.0687 (calcd m / z 463.0689 [M+2+H]) + ) Formula C 21 H 21 BrN2O5.

[0105] IR (KBr) v max3427, 2925, 2851, 2144, 1596, 1514, 1384, 1249, 1035, 866, 754, 695, 496 cm -1 .

[0106] UV λ max = 240 nm, 290 nm.

[0107] The single crystal of the indolylethylamine derivative prepared in this example was measured by X-ray single crystal diffraction method, and the crystallographic data was stored in the Cambridge Crystallographic Data Center (CCDC) with the storage number of CCDC 2288643.

[0108] Example 6

[0109] The clinically used lipid-lowering therapeutic drug fenofibrate was used as a positive control drug, the corresponding solvent was used as a negative control, oleic acid (OA) induction was used as an experimental group, and mouse normal liver cells AML12 were used as a test cell strain.

[0110] (1) Cell culture: AML12 cells were cultured in DMEM / F12 medium containing 10% inactivated fetal bovine serum FBS, 1 μg / ml recombinant human insulin, 0.55 μg / ml human transferrin, 0.67 ng / ml sodium selenite, 1 mM dexamethasone, 100 U / ml penicillin and 0.1 mg / ml streptomycin at 37°C in a 5% CO2 environment.

[0111] According to the results of the pre-experiment cell growth rate, 190 μL of cells with a certain density were inoculated in a 96-well culture plate (about 2×10 3 ~ 4×10 3 cells per well), and the cells were cultured in a saturated humidity, 5% CO2, 37°C environment for 24 h. After 24 h of culture, 10 μL of indolylethylamine derivative with a final concentration of 5 μM, 10 μM and 20 μM was added to the test wells, respectively, and 5 replicate wells were set for each sample. The negative control wells were added with 10 μL of blank matrix, and incubated for 24 h.

[0112] According to the results of the pre-experiment cell growth rate, 2 mL of cells with a certain density were inoculated in a 6-well culture plate (about 1×10 5 ~ 2×10 5The cells were incubated for 24 h under the condition of saturated humidity, 5% CO2, and 37°C. After 24 h of incubation, the old medium was removed, and 2 mL of medium containing 20 μM fenofibrate was added to the positive control wells. 2 mL of medium containing 5 μM, 10 μM, and 20 μM of the indolylethanamine derivative was added to the test wells, respectively. The cells were incubated for 2 h. After 2 h of incubation, 500 μM of oleic acid (OA) was added to all wells except the negative control wells, and the same volume of blank medium was added to the negative control wells. The cells were incubated for another 24 h.

[0113] (2) Drug toxicity test: According to experiment (1), after the cells in the 96-well plate were incubated for 24 h, the medium was removed, 100 μL of medium and 10 μL of CCK-8 reagent were added to each well, and the cells were incubated for another 1 h. The OD value was measured at a measuring wavelength of 450 nm and a reference wavelength of 620 nm by using an enzyme-labeled instrument. The cell proliferation inhibition rate of each dosing well was calculated, and the results are shown in A of Figure 2 As shown in A of Figure 2 , when the dosing concentration of the four indolylethanamine derivatives (compound 4, compound 5, compound 8, and compound 9) was 20 μM, the cells had no obvious toxicity except that compound 5 had slight cytotoxicity. As shown in C of , when the dosing concentration of compound 9 was 20 μM or less, the cells had no obvious toxicity.

[0114] Figure 2 (3) Triglyceride (TG) content detection: According to experiment (1), after the cells in the 6-well plate were incubated for 24 h, the medium was removed, and the cells were washed twice with PBS. The TG content of each well was measured by using a TG detection kit (Nanjing Jiancheng, A110-1-1), and the results are shown in B of Figure 2 As shown in B of , 20 μM of compound 4, compound 5, compound 8, and compound 9 can reduce the TG level of OA-induced cells. As shown in D of

[0115] , 5, 10, and 20 μM of compound 9 can dose-dependently reduce the TG level of OA-induced cells. Figure 3 (4) Oil red O staining detection: According to experiment (1), after the cells in the 6-well plate were incubated for 24 h, the medium was removed, and the cells were washed twice with PBS. According to the oil red O kit instruction, the cell lipid droplet staining was performed, and the cell nucleus was counterstained with hematoxylin for 10 min. The staining condition was observed and photographed by using an inverted microscope, and the results are shown in

[0116] As shown in , compared with the negative control group, the number and volume of cell lipid droplets increased significantly after OA acted on the AML12 cells for 24 h. Compared with the experimental group, the number and volume of cell lipid droplets significantly decreased after 5, 10, and 20 μM of compound 9 was added. These results suggest that compound 9 can effectively inhibit the generation of cell lipids induced by OA and reduce the level of cell lipids.(5) Bodipy lipid droplet fluorescence detection: Prepare 2 mg / mL Bodipy staining solution in PBS. According to experiment (1), after 6-well plate cultured cells were treated with drug for 24 h, rinse the cells quickly with 3 mL PBS to remove the culture medium. Incubate the cells in Bodipy staining solution at 37 °C for 30 min in the dark. Rinse the cells quickly with 3 mL PBS to remove the staining solution. Use fluorescence microscope with Ex / Em = 488 / 510 nm (FITC filter) to measure the fluorescence. The results are shown in Figure 4 Figure 6. Compared with the negative control group, the number and volume of cell lipid droplets increased significantly after OA acted on AML12 cells for 24 h. Compared with the experimental group, the number and volume of cell lipid droplets decreased significantly after 5, 10 and 20 μΜ compound 9 was administered. These results suggest that compound 9 can effectively inhibit the generation of cell lipids induced by OA and reduce the level of cell lipids.

[0117] (6) PCR: According to experiment (1), after 6-well plate cultured cells were treated with drug for 24 h, use SteadyPure RNA extraction kit (Accurate Biology, AG21024) to extract RNA. Use MonScript TM RTIII All-in-One Mix with dsDNase (Monad, MR05101S) to synthesize the corresponding cDNA. Use MonAmp TM Green qPCR Mix (Monad, MQ10201S) to detect the expression level of genes by qRT-PCR, and use 2 -ΔΔCt method to calculate the relative expression of target gene mRNA. The detected genes are as follows: ATGL, CPT1a, HSL, PPARa, VLDLR, β-actin. The results are shown in Figure 5 Figure 7. 5, 10 and 20 μΜ compound 9 can dose-dependently increase the mRNA expression level of PPARa, CPT1a, HSL, ATGL and VLDLR in OA-induced cells.

[0118] (7) Western Blot: According to experiment (1), after 24h of drug treatment, the culture medium of 6-well plate cultured cells was aspirated, and the cells were washed twice with PBS, and then the cells were homogenized in RIPA buffer containing protease inhibitors. The supernatant was separated by centrifugation at 12000xg for 5min, the protein content was determined, and the sample was frozen at -80℃ for use. Cell lysates were separated by 10% SDS-PAGE and transferred to a polyvinylidene fluoride membrane. SDS-PAGE was performed, and the proteins were detected using their respective antibodies. The bound antibodies were visualized by chemiluminescence. The following primary antibodies were used: ACC (Santa, sc-137104), p-ACC (Santa, sc-271965), HSL (Santa, sc-74489), PPARa (Santa, sc-398394), VLDLR (Santa, sc-18824), CPT1a (Abeam, ab234111), β-actin (Abclonal, ac026). The results are shown in Figures 1-4. Figure 6 As shown in A and B in the figure, 5, 10 and 20μM compound 9 can dose-dependently increase the protein expression levels of PPARa, CPT1a, HSL, VLDLR and ACC phosphorylation in OA-induced cells.

[0119] In summary, the indole ethylamine derivative prepared in the present application can reduce the lipid accumulation in hepatocytes induced by unsaturated fatty acid oleic acid.

[0120] The above-described embodiments of the present application are described in detail, and it should be understood that the above-described is only a specific embodiment of the present application, and is not used to limit the present application, and any modification, supplement or similar way of substitution made within the principle range of the present application should be included in the protection scope of the present application.

Claims

1. An indoleethylamine derivative, characterized in that, The structural formula is shown in formula (Ⅰ) or formula (Ⅱ): In formula (I), R1 is a hydroxyl or methoxy-substituted amino group; in formula (II), X is bromine; R2 is any of the following formulas. To replace the position; 2. The method for preparing the indoleethylamine derivative according to claim 1, characterized in that, When the indoleethylamine derivative has the structural formula shown in formula (I), the preparation method includes: In a solution of tryptamine, triethylamine and di-tert-butyl dicarbonate were added sequentially to yield compound 1. Compound 1 of SO2 reacts with the oxidant 2,3-dichloro-5,6-dicyanobenzoquinone to give compound 2. SO3 adds sodium hydride to a solution of compound 2, followed by the addition of p-methoxybenzyl halide, and the reaction yields compound 3. Compound 3 of SO4 and diisobutylaluminum hydride react in an organic solvent to give an indoleethylamine derivative with a hydroxyl group at R1. And / or, by sequentially adding methoxyamine hydrochloride and triethylamine to an organic solution of an indoleethylamine derivative in which R1 is a hydroxyl group, the reaction yields an indoleethylamine derivative in which R1 is a methoxy-substituted amino group.

3. The method for preparing the indoleethylamine derivative according to claim 2, characterized in that, In step S01, the reaction temperature is 10-50℃ and the reaction time is 1-5h.

4. The method for preparing the indoleethylamine derivative according to claim 2, characterized in that, In step S02, tetrahydrofuran and water are used as solvents in the reaction system, the reaction temperature is -10 to 60°C, and the reaction time is 1 to 6 hours.

5. The method for preparing the indoleethylamine derivative according to claim 2, characterized in that, In step S03, the reaction temperature is -10 to 50°C, the reaction time is 0.5 to 5 hours, and the p-methoxybenzyl halide is 4-methoxybenzyl bromide.

6. The method for preparing the indoleethylamine derivative according to claim 2, characterized in that, In step S04, the reaction temperature of compound 3 and diisobutylaluminum hydride is -100 to 20°C, and the reaction time is 0.5 to 3 hours. And / or, the reaction temperature of indoleethylamine derivatives with hydroxyl groups (R1) with methoxyamine hydrochloride and triethylamine is 80-150℃, and the reaction time is 15-30h.

7. The method for preparing the indoleethylamine derivative according to claim 1, characterized in that, When the indoleethylamine derivative has the structural formula shown in formula (II), the preparation method includes: S11: Sodium hydride was mixed with a solution of 6-bromoindole, stirred, and then p-methoxybenzyl halide was added to give compound 6. In the presence of aluminum trichloride, compound 6 reacts with methylmalonyl chloride to yield compound 7. Adding p-acetaminobenzenesulfonyl azide and triethylamine to a solution of compound 7 yields an indoleethylamine derivative R2 as shown in formula (III). And / or, R2 is an indoleethylamine derivative as shown in formula (III), or R1 is an indoleethylamine derivative with an amino group substituted with a methoxy group. The reaction in trifluorotoluene yields intermediate 1. 2,6-Dimethylaniline and trimethylsilyl trifluoromethanesulfonate are added to a dichloromethane solution of intermediate 1, and the reaction yields R2, an indoleethylamine derivative as shown in formula (IV).

8. The method for preparing the indoleethylamine derivative according to claim 7, characterized in that, In step S11, the p-methoxybenzyl halide is 4-methoxybenzyl bromide, the reaction temperature is -20 to 60°C, and the reaction time is 3 to 9 hours.

9. The method for preparing the indoleethylamine derivative according to claim 7, characterized in that, In step S12, the reaction temperature of compound 6 and methylmalonyl chloride is -20 to 50°C, and the reaction time is 8 to 15 h; the reaction temperature of compound 7 with p-acetaminobenzenesulfonyl azide and triethylamine is -20 to 60°C, and the reaction time is 6 to 18 h. And / or, the reaction temperature of R2 as an indoleethylamine derivative of formula (III) and the reaction time of R1 as an indoleethylamine derivative of a methoxy-substituted amino group in trifluorotoluene are 50-150℃ and 2-8h; the reaction temperature of intermediate 1 with 2,6-dimethylaniline and trimethylsilyl trifluoromethanesulfonate are -20-40℃ and 1-5h.

10. A drug, characterized in that, Including the indoleethylamine derivative of claim 1, the drug is used for anti-lipid accumulation, anti-oxidation and / or anti-inflammation.

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