A tryptanthrin compound, a preparation method and application thereof

Abstract

The application belongs to the technical field of organic compound preparation, and discloses a tryptanthrin compound and a preparation method and application thereof. The preparation method of the tryptanthrin compound is that an indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazin-2,4(1H)-dione, triethylamine and toluene are reacted to obtain the tryptanthrin compound. The tryptanthrin compound obtained by the method has a novel structure, shows an anti-ulcerative colitis activity, and has a potential to be developed into an anti-ulcerative colitis drug.

Description

Technical Field

[0001] This invention relates to the field of organic compound preparation technology, and in particular to a tryptamine ketone compound, its preparation method and application. Background Technology

[0002] Ulcerative colitis (UC) is a chronic, nonspecific inflammatory bowel disease that primarily affects the mucosa and submucosa of the colon and rectum. Typical symptoms include recurrent diarrhea, bloody and mucous stools, abdominal pain, tenesmus (a feeling of incomplete bowel movement), and weight loss. Severe cases may be accompanied by fever, anemia, and other systemic symptoms. The lesions typically begin in the rectum and extend continuously towards the proximal colon; the extent of inflammation correlates with disease severity. The etiology is not fully understood, but it is likely related to the interaction of multiple factors, including genetic susceptibility, abnormal activation of the immune system, gut microbiota dysbiosis, and environmental factors such as diet and smoking. Treatment aims to control inflammation, relieve symptoms, and reduce recurrence. Mild to moderate cases are commonly treated with aminosalicylic acid derivatives (such as mesalazine), while moderate to severe or refractory cases require treatment with corticosteroids, immunosuppressants (such as azathioprine), or biologics (such as anti-TNF-α monoclonal antibodies). Developing new drugs for treating ulcerative colitis is a pressing challenge in this field.

[0003] Tryptanthrin is a natural indole alkaloid widely found in medicinal plants such as Isatis indigotica. In recent years, it has become a research hotspot for the treatment of ulcerative colitis due to its significant anti-inflammatory and immunomodulatory activities. Studies have shown that tryptanthrin can inhibit intestinal inflammatory responses through multiple targets: (1) blocking key signaling pathways such as NF-κB and STAT3, and reducing the expression of pro-inflammatory factors such as TNF-α, IL-6, and IL-1β; (2) regulating the Th17 / Treg cell balance and inhibiting excessive immune responses; and (3) enhancing intestinal barrier function and reducing oxidative stress damage. Animal experiments have shown that tryptanthrin can significantly alleviate the symptoms of colitis induced by sodium dextran sulfate (DSS) in mice, and reduce the disease activity index and histopathological score. To improve efficacy and reduce toxicity, researchers have modified the structure of tryptanthrin and developed a series of derivatives. For example, the introduction of sulfonic acid groups can enhance water solubility, fluorinated derivatives show stronger NF-κB inhibitory activity, and C-6 hydroxyl-substituted derivatives show better antioxidant capacity. Currently, tryptophan offers a potential direction for the development of drugs for ulcerative colitis. Developing anti-ulcerative colitis drugs based on tryptophan is of great significance for the development of novel drug treatments.

[0004] Therefore, the discovery of a novel tryptophanone compound with anti-ulcerative colitis activity, along with its preparation method and applications, is of great significance. Summary of the Invention

[0005] In view of this, the present invention provides a tryptamine ketone compound, its preparation method and application, the purpose of which is to solve the problems of low efficacy and high toxicity of existing tryptamine ketone compounds.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] This invention provides a tryptamine ketone compound, the structural formula of which is shown below:

[0008]

[0009] R1, R2, and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso, or trifluoromethoxy.

[0010] Preferably, the structural formula of the tryptamine ketone compound includes:

[0011]

[0012]

[0013] The present invention also provides a method for preparing the aforementioned tryptamine ketone compound, comprising the following steps:

[0014] The indole-2,3-dione derivative, 2H-thiopheno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene were reacted to give tryptamine ketone compounds.

[0015] Preferably, the structural formula of the indole-2,3-dione derivative is:

[0016]

[0017] R1, R2, and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso, or trifluoromethoxy.

[0018] Preferably, the ratio of the indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene is 0.8–1.2 mmol: 0.8–1.2 mmol: 0.8–1.2 mL: 8–12 mL.

[0019] Preferably, the reaction time is 4 to 8 hours and the reaction temperature is 80 to 100°C.

[0020] The present invention also provides the application of the aforementioned tryptamine ketone compound in the preparation of anti-inflammatory drugs.

[0021] As can be seen from the above technical solution, compared with the prior art, the present invention has the following beneficial effects:

[0022] This invention prepares a tryptamine ketone compound with a novel structure, which exhibits anti-ulcerative colitis activity and has the potential to be developed into an anti-ulcerative colitis drug. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0024] Figure 1 The graph shows the NO secretion induced by bacterial lipopolysaccharide by the compounds obtained in Examples 1-16;

[0025] Figure 2 The graph shows the effect of compound 2l obtained in Example 12 on cell viability;

[0026] Figure 3 The graph shows the effect of compound 2l obtained in Example 12 on the disease activity index;

[0027] Figure 4 The effect of compound 2l obtained in Example 12 on the body weight of mice with ulcerative colitis;

[0028] Figure 5 The effect of compound 2l obtained in Example 12 on colon length in mice with ulcerative colitis;

[0029] Figure 6 This is a statistical graph showing the effect of compound 2l obtained in Example 12 on the colon length of mice with ulcerative colitis. Detailed Implementation

[0030] This invention provides a tryptamine ketone compound, the structural formula of which is shown below:

[0031]

[0032] R1, R2, and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso, or trifluoromethoxy.

[0033] In this invention, the structural formula of tryptophan ketone in the tryptophan ketone compound is as follows:

[0034]

[0035] In this invention, the preferred structural formula of the tryptamine ketone compound includes:

[0036]

[0037]

[0038] The present invention also provides a method for preparing the aforementioned tryptamine ketone compound, comprising the following steps:

[0039] The indole-2,3-dione derivative, 2H-thiopheno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene were reacted to give tryptamine ketone compounds.

[0040] In this invention, the preferred structural formula of the indole-2,3-dione derivative is:

[0041]

[0042] The R1, R2, and R3 are preferably hydrogen, halogen, methoxy, methyl, nitroso, or trifluoromethoxy.

[0043] In this invention, the preferred ratio of the indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine, and toluene is 0.8–1.2 mmol: 0.8–1.2 mmol: 0.8–1.2 mL: 8–12 mL, more preferably 0.9–1.1 mmol: 0.9–1.1 mmol: 0.9–1.1 mL: 9–11 mL, and even more preferably 1 mmol: 1 mmol: 1 mL: 10 mL.

[0044] In this invention, the reaction is a condensation reaction, and the synthetic route of the reaction is as follows:

[0045]

[0046] In this invention, the reaction time is preferably 4 to 8 hours, more preferably 5 to 7 hours, and even more preferably 6 hours; the reaction temperature is preferably 80 to 100°C, more preferably 85 to 95°C, and even more preferably 90 to 92°C.

[0047] In this invention, the reaction is followed by sequential cooling, filtration, washing, and purification;

[0048] The cooling temperature is preferably 20-30°C, more preferably 22-28°C, and even more preferably 25-26°C;

[0049] The washing reagent is preferably methanol, and the washing is preferably performed 3 to 6 times;

[0050] The purification is preferably carried out by silica gel column chromatography, wherein the eluent for the silica gel column chromatography is a hexane-ethyl acetate system, and the volume ratio of hexane to ethyl acetate is preferably 3 to 5:1, more preferably 4:1.

[0051] The present invention also provides the application of the aforementioned tryptamine ketone compound in the preparation of anti-inflammatory drugs.

[0052] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0053] In the embodiments, the structural formula of the indole-2,3-dione derivative is as follows:

[0054]

[0055] R1, R2, and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso, or trifluoromethoxy.

[0056] Example 1

[0057] Compound 2a:

[0058]

[0059] Preparation of compound 2a:

[0060] 1 mmol of indole-2,3-dione (R1, R2 and R3 are all hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0061] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2a. The yield was calculated to be 70%.

[0062] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.43(d,J=8.0Hz,1H),8.33(d,J=5.2Hz,1H),7.89–7.80(m,2H),7.64(d,J=5.2Hz,1H),7.47(t,J=7.5Hz,1H)ppm; 13C NMR (101MHz, DMSO-d6) δ182.11,155.50,154.23,147.08,145.97,137.65,136.62,127.15,126.66,126.42,124.91,122.51,117.11ppm.

[0063] Example 2

[0064] Compound 2b:

[0065]

[0066] Preparation of compound 2b:

[0067] 1 mmol of 5-fluoroindole-2,3-dione (R2 is fluorine, and R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0068] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2b. The yield was calculated to be 56%.

[0069] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.44 (dd, J=8.8, 4.2Hz, 1H), 8.34 (d, J=5.2Hz, 1H), 7.7 8(dd,J=7.1,2.8Hz,1H),7.69(td,J=9.0,2.8Hz,1H),7.65(d,J=5.3Hz,1H)ppm; 13 C NMR(101MHz,DMSO-d6)δ181.22,159.18,155.43,154.05,142.26,136.76,12 6.66,126.37,124.39,123.81,123.57,118.90,118.82,112.00,111.76ppm.

[0070] Example 3

[0071] Compound 2c:

[0072]

[0073] Preparation of compound 2c:

[0074] 1 mmol of 5-chloroindole-2,3-dione (R2 is chlorine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0075] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2c. The yield was calculated to be 69%.

[0076] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.39(d,J=8.5Hz,1H),8.32(d,J=5.2Hz,1H),7.92(d,J=2.3Hz,1H),7.86(dd,J=8.5,2.3Hz,1H),7.63(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ181.00,155.53,154.13,147.05,144.44,136.97,136.73,131.48,126.74,126.31,124.52,124.34,118.70ppm.

[0077] Example 4

[0078] Compound 2d:

[0079]

[0080] Preparation of compound 2d:

[0081] 1 mmol of 5-bromoindole-2,3-dione (R2 is bromine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0082] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2d. The yield was calculated to be 85%.

[0083] Test results: 1H NMR (400MHz, DMSO-d6) δ8.39–8.33(m,2H),8.07–7.99(m,2H),7.66(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ180.84,155.51,154.07,146.81,144.77,139.57,136.93,127.27,126.70,126.27,124.53,119.37,118.98ppm.

[0084] Example 5

[0085] Compound 2e:

[0086]

[0087] Preparation of compound 2e:

[0088] 1 mmol of 5-iodoindole-2,3-dione (R2 is iodine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0089] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2e. The yield was calculated to be 81%.

[0090] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.35(d,J=5.3Hz,1H),8.22(d,J=8.3Hz,1H),8.19(d,J=1.8Hz,1H),8.16(t,J=2.3Hz,1H),7.65(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ180.82,155.53,154.08,146.52,145.43,145.23,136.86,132.75,126.68,126.28,124.51,119.09,91.71ppm.

[0091] Example 6

[0092] Compound 2f:

[0093]

[0094] Preparation of compound 2f:

[0095] 1 mmol of 6-chloroindole-2,3-dione (R3 is chlorine, R1 and R2 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0096] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2f. The yield was calculated to be 88%.

[0097] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.39(d,J=1.9Hz,1H),8.33(d,J=5.2Hz,1H),7.88(d,J=8.0Hz,1H),7.64(d,J=5.2Hz,1H),7.53(dd,J=8.1,1.9Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ181.33,155.98,154.80,147.57,147.12,141.87,137.56,127.63,127.20,126.88,126.54,121.97,117.43ppm.

[0098] Example 7

[0099] Compound 2g:

[0100]

[0101] Preparation of 2g of compound:

[0102] 1 mmol of 4-chloroindole-2,3-dione (R1 is chlorine, R2 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0103] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to obtain 2 g of a yellow solid compound. The yield was calculated to be 46%.

[0104] Test results:1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=8.0Hz,1H),8.41(d,J=5.3Hz,1H),7.88(t,J=8.1Hz,1H),7.72(d,J=5.2Hz,1H),7.57(d,J=8.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ179.22,155.52,154.13,147.18,146.67,138.36,137.00,131.48,128.29,126.69,126.15,119.39,115.83ppm.

[0105] Example 8

[0106] Compound 2h:

[0107]

[0108] Preparation of compound 2h:

[0109] 1 mmol of 6-bromoindole-2,3-dione (R3 is bromine, R1 and R2 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0110] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to obtain a yellow solid compound for 2 hours. The yield was calculated to be 52%.

[0111] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.59(d,J=1.7Hz,1H),8.37(d,J=5.2Hz,1H),7.82(d,J=8.1Hz,1H),7.71(dd,J=8.1,1.7Hz,1H),7.67(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ180.84,155.51,154.07,146.81,144.77,139.57,136.93,127.27,126.70,126.27,124.53,119.37,118.98ppm.

[0112] Example 9

[0113] Compound 2i:

[0114]

[0115] Preparation of compound 2i:

[0116] 1 mmol of 5-methylindole-2,3-dione (R2 is methyl, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0117] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to obtain a yellow solid compound 2i. The yield was calculated to be 85%.

[0118] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.32(d,J=5.3Hz,1H),8.29(d,J=8.1Hz,1H),7.68(d,J=1.8Hz,1H),7.66–7.61(m,2H),2.40(s,3H)ppm; 13 C NMR (101MHz, DMSO-d6) δ182.15,155.46,154.03,147.17,143.88,137.92,136.93,136.43,126.60,126.45,124.99,122.53,116.85,20.41ppm.

[0119] Example 10

[0120] Compound 2j:

[0121]

[0122] Preparation of compound 2j:

[0123] 1 mmol of 5-methoxyindole-2,3-dione (R2 is methoxy, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0124] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to obtain a yellow solid compound 2j. The yield was calculated to be 87%.

[0125] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.34–8.28(m,2H),7.62(d,J=5.3Hz,1H),7.41–7.36(m,2H),3.86(s,3H)ppm; 13 C NMR (101MHz, DMSO-d6) δ182.00,158.15,155.36,153.86,147.32,139.71,136.29,126.58,126.53,123.65,123.26,118.29,108.97,56.04ppm.

[0126] Example 11

[0127] Compound 2k:

[0128]

[0129] Preparation of compound 2k:

[0130] 1 mmol of 5-trifluoromethoxyindole-2,3-dione (R2 is trifluoromethoxy, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0131] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2k. The yield was calculated to be 65%.

[0132] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=8.7Hz,1H),8.33(d,J=5.2Hz,1H),7.89(d,J=2.5Hz,1H),7.84(dd,J=8.6,2.6Hz,1H),7.64(d,J=5.2Hz,1H)ppm; 13C NMR (101MHz, DMSO-d6) δ180.87,155.46,154.08,17.23,146.44,144.44,136.99,129.90,126.70,126.23,124.28,121.28,118.74,117.84ppm.

[0133] Example 12

[0134] Compound 2l:

[0135]

[0136] Preparation of compound 2l:

[0137] 1 mmol of 5-nitroindole-2,3-dione (R2 is nitroso, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0138] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2l. The yield was calculated to be 61%.

[0139] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.65(dd,J=8.8,2.4Hz,1H),8.58(d,J=8.8Hz,1H),8.48(d,J=2.3Hz,1H),8.35(d,J=5.2Hz,1H),7.65(d,J=5.2Hz,1H)ppm; 13 CNMR(101MHz,DMSO-d6)δ180.34,155.35,154.18,149.21,147.38,145.80,137.58,132.51,126.81,126.03,123.44,119.55,117.73ppm.

[0140] Example 13

[0141] Compound 2m:

[0142]

[0143] Preparation of compound 2m:

[0144] 1 mmol of 4,5-dichloroindole-2,3-dione (R1 and R2 are both chlorine, and R3 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0145] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2m. The yield was calculated to be 54%.

[0146] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.57(s,1H),8.38(d,J=5.2Hz,1H),8.22(s,1H),7.68(d,J=5.3Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ179.89,155.49,153.99,146.82,144.41,138.88,137.28,129.84,126.71,126.36,125.93,122.92,118.53ppm.

[0147] Example 14

[0148] Compound 2n:

[0149]

[0150] Preparation of compound 2n:

[0151] 1 mmol of 5,6-dichloroindole-2,3-dione (R2 and R3 are both chlorine, and R1 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product.

[0152] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2n. The yield was calculated to be 67%.

[0153] Test results: 1H NMR (400MHz, DMSO-d6) δ8.46–8.34(m,2H),8.07(d,J=8.6Hz,1H),7.67(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ178.39,155.52,154.00,146.58,145.55,137.70,137.28,130.34,129.46,126.72,126.02,121.17,116.78ppm.

[0154] Example 15

[0155] Compound 2o:

[0156]

[0157] Preparation of compound 2o:

[0158] 1 mmol of 4-chloro-5-fluoroindole-2,3-dione (R1 is chlorine, R2 is fluorine, R3 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0159] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to obtain a yellow solid compound 2o. The yield was calculated to be 69%.

[0160] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.42(dd,J=8.8,3.8Hz,1H),8.36(d,J=5.3Hz,1H),7.86(t,J=9.2Hz,1H),7.67(d,J=5.2Hz,1H)ppm; 13 C NMR(101MHz,DMSO-d6)δ178.63,157.07,155.40,154.62,153.98,146.85,142.92,13 7.07,126.64,126.10,123.98,123.75,120.94,118.59,118.38,116.88,116.80ppm.

[0161] Example 16

[0162] Compound 2p:

[0163]

[0164] Preparation of compound 2p:

[0165] 1 mmol of 5-fluoro-6-chloroindole-2,3-dione (R2 is fluorine, R3 is chlorine, R1 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90 °C for 6 h to obtain the crude product;

[0166] The obtained product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (using a hexane-ethyl acetate eluent system with a volume ratio of hexane to ethyl acetate of 4:1) to give a yellow solid compound 2p. The yield was calculated to be 63%.

[0167] Test results: 1 H NMR (400MHz, DMSO-d6) δ8.50(d,J=6.1Hz,1H),8.34(d,J=5.2Hz,1H),8.04(d,J=7.5Hz,1H),7.64(d,J=5.2Hz,1H)ppm; 13 C NMR(101MHz,DMSO-d6)δ180.69,157.65,155.94,155.18,154.47,147.52,142.5 6,142.53,137.68,127.20,126.51,123.53,123.46,119.15,113.86,113.62ppm.

[0168] The NO secretion levels of the compounds obtained in Examples 1-16 were measured respectively:

[0169] RAW264.7 cells were diluted with complete culture medium to a concentration of 2.0 × 10⁶ cells per well. 5 Cells were seeded into 96-well plates. After cell attachment, the supernatant was gently aspirated, and 90 μL of complete culture medium solution of compounds 1–16 (5 μM concentration) was added to each well. After 2 h, 10 μL of bacterial lipopolysaccharide (LPS) at a concentration of 1 μg / mL was added to each well, and the cells were cultured for another 20 h before collecting the supernatant.

[0170] NO content was determined using the Griess method: 50 μL of cell supernatant was taken from each sample well, and 50 μL of sulfanilamide solution was added. The mixture was reacted in the dark for 10 min. Then, 50 μL of N-(1-naphthyl)ethylenediamine solution was added, and the reaction was continued in the dark for another 10 min. Finally, the absorbance of each well was measured at 562 nm using a microplate reader. The results are shown below. Figure 1 As shown.

[0171] The graphs showing the NO secretion induced by bacterial lipopolysaccharide in the compounds obtained in Examples 1-16 are shown below. Figure 1 As shown. By Figure 1 Compounds 2c, 2d, 2e, 2h, 2j, 2k, and 2l exhibited inhibitory activity against nitric oxide (relative to the inflammation model group, i.e., the LPS group in the figure), demonstrating that these compounds possess anti-inflammatory activity and have the potential to become anti-inflammatory drugs. In particular, 2l showed the most significant inhibition of NO secretion at the same molar concentration, exhibiting significant anti-inflammatory activity and potential for further development.

[0172] MTT assay was performed on compound 2l obtained in Example 12:

[0173] The specific steps are as follows: RAW264.7 cells were prepared with complete culture medium to a density of 1.25 × 10⁶ cells / year. 5 Single-cell suspensions of 100 μL / mL were seeded into each well of a 96-well plate. After culturing the cells under suitable conditions for 24 h, the original culture medium was discarded, and 100 μL of drug solutions of different concentrations were added. The test drugs were prepared with six concentration gradients: 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, and 3.125 μM (the drug stock solution was prepared with biological grade DMSO to 1 mmol / L, and then serially diluted with culture medium before use). A blank control group and a control group were also established. After culturing for another 20 h, 10 μL of LTT solution was added to each well, incubated for 4 h, and then DMSO was added, followed by shaking for 1 min. Finally, the absorbance (A) of each well at 490 nm was measured using a microplate reader, and cell viability was calculated according to the following formula. The results are shown below. Figure 2 As shown.

[0174] Cell viability (%) = (A-drug group - A-control group) / (A-control group - A-blank group) × 100%.

[0175] The following performance tests were performed on compound 2l obtained in Example 12:

[0176] Animal grouping and administration: Based on the results of MTT and NO secretion detection, tryptophan and compound 2l with good anti-inflammatory activity and low cytotoxicity were selected for the experiment.

[0177] The experimental animals were divided into the following groups: control group, model group, positive control group with added sulfasalazine (dose of 125 mg / kg), tryptophan group (dose of 78 mg / kg), and compound 2l group (dose of 108 mg / kg).

[0178] Model establishment: Sixty male SPF-grade C57BL / 6 mice, aged 6–8 weeks and weighing 18–22 g, were used in the experiment. The mice were acclimatized for one week before the experiment began. The control group mice drank purified water, while the other groups drank 2.5% dextran sulfate sodium solution to induce the formation of the UC mouse model.

[0179] Drug administration: The mice were administered the drug by gavage at the same time as the model was established. The control group and the model group were administered distilled water by gavage, while the sulfasalazine group, TRYP and compound 2l group were administered the corresponding dose of the drug by gavage. The gavage volume was 10 mL / kg, and the administration was repeated for 12 consecutive days.

[0180] Disease Activity Index (DAI) Score: From the first day of animal modeling, the mice's weight, stool consistency, and fecal bloodiness were recorded according to the DAI scoring table to calculate the Disease Activity Index (DAI). Test results are as follows: Figures 3-6 As shown.

[0181] The effect of compound 2l obtained in Example 12 on the histopathological score of colon tissue in UC mice is shown in the figure below. Figure 3 As shown. By Figure 3 It is evident that compound 2l has a better therapeutic effect than the positive control drug sulfasalazine.

[0182] The effect of compound 2l obtained in Example 12 on the body weight of mice with ulcerative colitis is as follows: Figure 4 As shown. By Figure 4 It is evident that, compared to the model group and the sulfasalazine group, mice treated with compound 2l had the highest average weight.

[0183] The effect of compound 2l obtained in Example 12 on colon length in mice with ulcerative colitis is as follows: Figure 5 and Figure 6 As shown. By Figure 5 and Figure 6 It is evident that, compared to the model group and the sulfasalazine group, mice treated with compound 2l had the longest colon.

[0184] Combination Figures 3-6 It is evident that compound 2l has therapeutic effects on ulcerative colitis and has the potential to be developed into a drug.

[0185] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A tryptophan ketone compound, characterized in that, The structural formulas of the tryptophan ketone compounds include: 。 2. The method for preparing a tryptamine ketone compound according to claim 1, characterized in that, Includes the following steps: The reaction of indole-2,3-dione derivatives, 2H-thienro[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene yields tryptamine ketone compounds; The structural formula of the indole-2,3-dione derivative is: , , , , , or .

3. The method for preparing a tryptamine ketone compound according to claim 2, characterized in that, The ratio of the indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine, and toluene is 0.8~1.2 mmol: 0.8~1.2 mmol: 0.8~1.2 mL: 8~12 mL.

4. The method for preparing a tryptamine ketone compound according to claim 2 or 3, characterized in that, The reaction time is 4-8 hours, and the reaction temperature is 80-100℃.

5. The use of the tryptamine ketone compound of claim 1 in the preparation of anti-inflammatory drugs.

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